EP3189048A1 - Aminoindane-, aminotetrahydronaphthalene- and aminobenzocyclobutane-derived prmt5-inhibitors - Google Patents

Abstract

A compound of formula (la), (lb) or (lc) wherein: n is 1 or 2; R^N is H or Me; R¹ is optionally one or more halo or methyl groups; R^2a and R^2b are independently selected from the group consisting of: (i) F; (ii) H; (iii) Me; and (iv) CH₂OH; R^2c and R^2d (if present) are independently selected from the group consisting of: (i) F; (ii) H; (iii) Me; and (iv) CH₂OH; R^3a and R^3b are independently selected from H and Me; R^4a is selected from OH, -NH₂, -C(=O)NH₂, and -CH₂OH; R^4b is either H or Me; R⁵ is either H or Me; A is either (i) optionally substituted phenyl; (ii) optionally substituted naphthyl; or (iii) optionally substituted C_5-12 heteroaryl.

Description

AMINOINDANE-, AMINOTETRAHYDRONAPHTHALENE- AND AMINOBENZOCYCLOBUTANE-DERIVED PRMT5-INHIBITORS

The present invention relates to amino-bicyclic compounds and their use as

pharmaceuticals, and in particular, in treating cancer and hemoglobinopathies.

Background to the Invention

Post-translational modification of proteins is a hallmark of signal transduction where cells are able to react quickly to changes or events in the cellular environment. Post-translational modification of proteins expands the structural and functional diversity of the proteome. The role of acetylation and phosphorylation of proteins has been extensively studied as highly reversible reactions for fine- tuning responses to external stimuli or changes in the environmental conditions. Recently, the importance of other types of protein modifications, including ubiquitination and methylation has begun to be recognized. The methylation of proteins and the enzymes that carry out these reactions has increased the dimensions of gene regulation by marking genes that are transcriptionally active or silenced. Protein methylation can occur on amino acids such as lysine, arginine, histidine, or proline, and on carboxy groups. Arginine methylation of mainly nuclear proteins is an important post-translational

modification process involved in structural remodelling of chromatin, signal transduction, cellular proliferation, nucleocytoplasmic shuttling, translation, gene transcription, DNA repair, RNA processing, or mRNA splicing. Methylation of proteins at arginine residues is catalysed by Protein Arginine

Methyltransferase enzymes. The Protein Arginine Methyl Transferase (PRMT) family of enzymes are evolutionarily conserved between organisms but differ in the number of members in different organisms. There are eleven members of the human PRMT family, eight of which have known enzymatic activity and target substrates. With the exception of PRMT2 and two recently identified putative PRMT genes (PRMT10 and PRMT1 1 ), all remaining proteins of the family possess enzymatic arginine methylation activity. PRMTs are subdivided into two types based on the methylation that they catalyse at the guanidinium group of arginine residues of substrate proteins. There are three nitrogens in the guanidinium group, potentially all of which could be methylated; the two Ψ-guanidino nitrogen atoms and the internal δ-guanidino nitrogen atom. Mono-methylation and dimethylation of arginine (MMA and DMA) is found in mammalian cells at one or both of the two Ψ-guanidino nitrogen atoms; dimethylation may be either symmetric or asymmetric. The third methylated arginine is generated by monomethylation of the internal δ-guanidino nitrogen atom of arginine and has so far been documented only in yeast proteins. Type I PRMT enzymes catalyse the formation of MMA and asymmetric dimethylarginine by di- methylating the same nitrogen atom of the guanidinium group, whereas Type II PRMT enzymes catalyse the formation of MMA and symmetric di-methylarginine by mono- methylating each of the terminal nitrogen atoms. Type III enzymes methylate the internal δ- guanidino nitrogen atom. Of the eight well characterised human PRMTs, PRMT1 , 3, 4, 6 and 8 are Type I enzymes, and PRMT5, 7 and 9 are Type II enzymes.

PRMTs catalyse the methylation of the guanidino nitrogen atoms of arginine residues through the transfer of a methyl group from S-adenosyl methionine (SAM). A by-product of the enzymatic methylation step is S-adenosyl-L-homocysteine (AdoHcy), which is hydrolyzed to adenosine and homocysteine by AdoHcy hydrolase (Krause et al., 2007).

PRMT5

PRMT5 (aka JBP1 , SKB1 , IBP72, SKBI his and HRMTIL5) is a Type II arginine

methyltransferase, and was first identified in a two-hybrid search for proteins interacting with the Janus tyrosine kinase (Jak2) (Pollack et al., 1999).

PRMT5 plays a significant role in control and modulation of gene transcription. Inter alia, PRMT5 is known to methylate histone H3 at Arg-8 (a site distinct from that methylated by

PRMT4) and histone H4 at Arg-3 (the same site methylated by PRMT1 ) as part of a complex with human SWI/SNF chromatin remodelling components BRG1 and BRM.

In addition to direct repressive histone marks induced by PRMT5, the enzyme's role in gene silencing is also mediated through the formation of multiprotein repressor complexes that include NuRD components, HDACs, MDB proteins and DNA methyltransferases, (Rank et al., 2010; Le Guezennec et al., 2006; Pal et al., 2003).

PRMT5 is involved in the methylation and functional modulation of the tumour suppressor protein p53. See (Berger, 2008; Durant et al., 2009; Jansson et al., 2008; Scoumanne et al., 2009). Most of the physiological functions of p53 are attributable to its role as a transcriptional activator, responding to agents that damage DNA. p53 status is wild type in approximately half of human cancer cases. These include 94% in cervix, 87% in blood malignancies, 85% in bones and endocrine glands, and 75% of primary breast cancer.

Restoration of p53 in cancer cells harbouring wild type p53, by way of inhibiting mechanisms that suppress its function leads to growth arrest and apoptosis, and is regarded as a potentially effective means of tumour suppression. p53 target genes have two alternative downstream effects: either they pause the cell cycle, allowing the DNA to be repaired, or, if repair is not possible, they activate processes leading to apoptosis (programmed cell death). How p53 'chooses' between these distinct outcomes is a central question in the field of tumour biology. p53 is replete with post-translational modifications. Phosphorylation was one of the first post- translational modifications to be clearly defined on p53. In the last decade it has become additionally clear that p53 is modified not only by phosphorylation, but that it is extensively modified by lysine acetylation and methylation, among other modifications. Indeed, besides histone proteins p53 is the most common protein substrate known for these post- translational modifications. However, despite the plethora of post-translational modifications, p53 has not been identified, until recently, as a substrate for arginine methylation.

Jansson et al (Jansson et al., 2008) discovered that PRMT5 is physically associated with a p53 cofactor called Strap. A co-factor complex that contains Strap et al binds to p53 in response to DNA damage. Jansson et al demonstrated that PRMT5 methylates p53 in vitro, and mapped the sites of methylation (R333, R335 and R337). They developed an antibody that specifically detects p53 methylated on these sites and confirmed that p53 is methylated in vivo. Jansson et al went on to show that p53 methylation requires PRMT5 and is increased in response to etoposide, a DNA damaging agent.

The role of PRMT5 and p53 arginine methylation on cell cycle regulation and DNA damage response have been explored by both Jansson et al and Scoumanne et al (Jansson et al., 2008; Scoumanne et al., 2009). Although some differences are evident between the results from the two groups in respect of cell cycle regulation in unperturbed cells (which may be ascribed to cell type specific effects and/or the actual nature of the experimental

arrangements), both groups report similar results with respect to the DNA damage response. In response to DNA damage, caused by a variety of agents, including doxorubicin, camptothecin and UV light, and also in response to treatment with Nutlin-3, knockdown of PRMT5 results in an increase in sub-G1 population and concomitant reduction in G1 cells and, in the presence of p53, a significant increase in apoptosis. Knockdown of PRMT5 also resulted in a reduced level of p21 , a key p53 target gene that regulates cell cycle arrest during the p53 response and MDM2, a p53 E3 ubiquitin ligase, but not PUMA, NOXA, AIP1 & APAF1 , p53 target genes linked to apoptosis.

Knockdown of PRMT5 (but not PRMT1 or CARM1/PRMT4) results in decreased p53 stabilisation, decreased basal p53 levels, and decreased p53 oligomerisation, and also decreased expression of elF4E a major component of translational machinery involved in ribosome binding to mRNA. Indeed, elF4E is a potent oncogene, which has been shown to promote malignant transformation in vitro and human cancer formation. Knockdown of PRMT5 would be expected to lead to a reduction in the level of arginine methylated p53. Consistent with arginine methylation status of p53 influencing the p53 response (reduced arginine methylation biasing the response to proapoptotic), Jannson et al showed that a p53 mutant in which each of the three critical arginine residues were substituted with lysine (p53KKK) retained the ability to induce apoptosis but its cell cycle arrest activity was significantly compromised.

Moreover, pS3KKK also has a significantly reduced ability to induce transcription of p21 , by contrast with APAF1 . The promoter binding specificity of wild-type p53 to key target genes is also significantly affected by arginine methylating status: Knockdown of PRMT5 results in decreased p53 binding to the promoter regions of the p21 and (intriguingly) PUMA genes, but does not affect p53 binding to the promoter regions of NOXA or APAF1 .

Taken together, it would seem that PRMT5 is a pro-survival factor, which regulates cell proliferation in unstressed conditions and modulates the p53 response during DNA damage. In particular, knockdown of PRMT5, leading to a reduction in the levels of arginine methylated p53, appears to bias the p53 DNA damage response to proapoptotic as opposed to cell cycle arrest.

PRMT5 is further linked to cancers in that it is aberrantly expressed in around half of human cancer cases. PRMT5 overexpression has been observed in patient tissue samples and cell lines of Prostate cancer (Gu et al., 2012), Lung cancer (Zhongping et al., 2012), Melanoma cancer (Nicholas et al., 2012), Breast cancer (Powers et al., 201 1 ), Colorectal cancer (Cho et al., 2012), Gastric cancer (Kim et al., 2005), Esophagus and Lung carcinoma (Aggarwal et al., 2010) and B-Cell lymphomas and leukemia (Wang, 2008). Moreover, elevated expression of PRMT5 in Melanoma, Breast and Colorectal cancers has been demonstrated to correlate with a poor prognosis.

Lymphoid malignancies including CLL are associated with over-expression of PRMT5.

PRMT5 is over-expressed (at the protein level) in the nucleus and cytosol in a number of patient derived Burkitt's lymphoma; mantle cell lymphoma (MCL); in vitro EBV-transformed lymphoma; leukaemia cell lines; and B-CLL cell lines, relative to normal CD19+ B

lymphocytes (Pal et al., 2007; Wang et al., 2008). Intriguingly, despite elevated levels of PRMT5 protein in these tumour cells, the levels of PRMT5 mRNA are reduced (by a factor of 2 - 5). Translation of PRMT5 mRNA is however, enhanced in lymphoma cells, resulting in increased levels of PRMT5 (Pal et al., 2007; Wang et al., 2008).

In addition to genomic changes, CLL, like almost all cancers, has aberrant epigenetic abnormalities characterised by global hypomethylation and hot-spots of repressive hypermethylation of promoters including tumour suppressor genes. While the role of epigenetics in the origin and progression of CLL remains unclear, epigenetic changes appear to occur early in the disease and specific patterns of DNA methylation are associated with worse prognosis (Chen et al., 2009; Kanduri et al., 2010). Global symmetric

methylation of histones H3R8 and H4R3 is increased in transformed lymphoid cell lines and MCL clinical samples (Pal et al., 2007), correlating with the overexpression of PRMT5 observed in a wide variety of lymphoid cancer cell lines and MCL clinical samples.

PRMT5 is therefore a target for the identification of novel cancer therapeutics.

PRMT5 Function and Hemoglobinopathies

Hemoglobin is a major protein in red blood cells and is essential for the transport of oxygen from the lungs to the tissues. In adult humans, the most common hemoglobin type is a tetramer called hemoglobin A, consisting of two a and two β subunits. In human infants, the hemoglobin molecule is made up of two a and two γ chains. The gamma chains are gradually replaced by subunits as the infant grows. The developmental switch in human β- like globin gene subtype from foetal (γ) to adult (β) that begins at birth heralds the onset of the hemoglobinopathies β-thalassemia and sickle cell disease (SCD). In β-thalassemia the adult chains are not produced. In SCD a point mutation in the coding sequence in the β globin gene leads to the production of a protein with altered polymerisation properties. The observation that increased adult γ-globin gene expression (in the setting of hereditary persistence of foetal hemoglobin (HPFH) mutations) significantly ameliorates the clinical severity of β-thalassemia and SCD has prompted the search for therapeutic strategies to reverse γ-globin gene silencing. To date, this has been achieved through pharmacological induction, using compounds that broadly influence epigenetic modifications, including DNA methylation and histone deacetylation. The development of more targeted therapies is dependent on the identification of the molecular mechanisms underpinning foetal globin gene silencing. These mechanisms have remained elusive, despite exhaustive study of the HPFH mutations, and considerable progress in many other aspects of globin gene regulation.

PRMT5 plays a critical role in triggering coordinated repressive epigenetic events that initiate with dimethylation of histone H4 Arginine 3 (H4R3me2s), and culminate in DNA methylation and transcriptional silencing of the γ-genes (Rank et al., 2010). Integral to the synchronous establishment of the repressive markers is the assembly of a PRMT5-dependent complex containing the DNA methyltransferase DNMT3A, and other repressor proteins (Rank et al., 2010). DNMT3A is directly recruited to bind to the PRMT5-induced H4R3me2s mark, and loss of this mark through shRNA-mediated knock-down of PRMT5, or enforced expression of a mutant form of PRMT5 lacking methyltransferase activity leads to marked upregulation of γ-gene expression, and complete abrogation of DNA methylation at the γ-promoter.

Treatment of human erythroid progenitors with non-specific methyltransferase inhibitors (Adox and MTA) also resulted in upregulation of γ-gene expression (He Y, 2013). Inhibitors of PRMT5 thus have potential as therapeutics for hemoglobinopathies such as β- thalassemia and Sickle Cell Disease (SCD).

The present inventors have developed particular substituted β-hydroxy amides inhibit the activity of PRMT5 and therefore may be of use in treating conditions ameliorated by the inhibition of the activity of PRMT5.

Summary of the Invention

A first aspect of the present invention provides a compound of formula la, lb or lc:

wherein:

n is 1 or 2;

R^N is H or Me;

R¹ is optionally one or more halo or methyl groups;

R^2a and R^2b are independently selected from the group consisting of:

(i) F:

(ii) H;

(iii) Me; and

(iv) CH₂OH;

R^2c and R^2d (if present) are independently selected from the group consisting of:

(i) F:

(ii) H;

(iii) Me; and

R^3a and R^3b are independently selected from H and Me;

R^4a is selected from OH, -NH₂, -C(=0)NH₂, and -CH2OH;

R^4b is either H or Me;

R⁵ is either H or Me;

A is either

(i) optionally substituted phenyl;

(ii) optionally substituted naphthyl; or (iii) optionally substituted C5-12 heteroaryl.

A second aspect of the present invention provides a compound of the first aspect for use in a method of therapy. The second aspect also provides a pharmaceutical composition comprising a compound of the first aspect and a pharmaceutically acceptable excipient.

A third aspect of the present invention provides a method of treatment of cancer, comprising administering to a patient in need of treatment, a compound of the first aspect of the invention or a pharmaceutical composition of the first aspect of the invention. The third aspect of the present invention also provides the use of a compound of the first aspect of the invention in the manufacture of a medicament for treating cancer, and a compound of the first aspect of the invention or pharmaceutical composition thereof for use in the treatment of cancer. As described below, the compound of the first aspect may be administered simultaneously or sequentially with radiotherapy and/or chemotherapy in the treatment of cancer.

A fourth aspect of the present invention provides a method of treatment of

hemoglobinopathies, comprising administering to a patient in need of treatment, a compound of the first aspect of the invention or a pharmaceutical composition of the first aspect of the invention. The fourth aspect of the present invention also provides the use of a compound of the first aspect of the invention in the manufacture of a medicament for treating hemoglobinopathies, and a compound of the first aspect of the invention or pharmaceutical composition of the first aspect of the invention for use in the treatment of

hemoglobinopathies.

Definitions

Cs-12 heteroaryl: The term "C5-12 heteroaryl" as used herein, pertains to a monovalent moiety obtained by removing a hydrogen atom from an aromatic structure having from 5 to 12 rings atoms, of which from 1 to 3 are ring heteroatoms. The term 'aromatic structure' is used to denote a single ring or fused ring systems having aromatic properties, and the term 'ring heteroatom' refers to a nitrogen, oxygen or sulphur atom.

In this context, the prefixes (e.g. C5-12, C5-6, etc.) denote the number of atoms making up the aromatic structure, or range of number of atoms making up the aromatic structure, whether carbon atoms or heteroatoms. Examples of C5-12 heteroaryl structures include, but are not limited to, those derived from: i : pyrrole (azole) (C5), pyridine (azine) (Ce); pyridone (Ce); indole (Cg); quinoline (C10);

O1 : furan (oxole) (C5);

Si : thiophene (thiole) (C5);

N1O1 : oxazole (C5), isoxazole (C5), isoxazine (Ce);

N2O1 : oxadiazole (furazan) (C5);

N1S1 : thiazole (C5), isothiazole (C5);

N2S1 : thiadiazole (C₅)

N₂: imidazole (1 ,3-diazole) (C5), pyrazole (1 ,2-diazole) (C5), pyridazine (1 ,2-diazine) (Ce), pyrimidine (1 ,3-diazine) (Ce) (e.g., cytosine, thymine, uracil), pyrazine (1 ,4-diazine) (Ce); benzimidazole (Cg)

N3: triazole (C5), triazine (Ce). Optional substituents

The optional substituents for the phenyl, naphthyl and C5-12 heteroaryl groups in A may be selected from the following groups.

C1-4 alkyl: The term "C1-4 alkyl" as used herein, pertains to a monovalent moiety obtained by removing a hydrogen atom from a carbon atom of a saturated hydrocarbon compound having from 1 to 4 carbon atoms.

Examples of saturated alkyl groups include, but are not limited to, methyl (Ci ), ethyl (C2), propyl (C3), and butyl (C₄).

Examples of saturated linear alkyl groups include, but are not limited to, methyl (Ci ), ethyl (C2), n-propyl (C3), and n-butyl (C.₍).

Examples of saturated branched alkyl groups include iso-propyl (C3), iso-butyl (C.₍), sec-butyl (C₄) and tert-butyl (C₄).

C1-1 fluoroalkyl: The term "C-M fluoroalkyl" refers to a Ci-₄ alkyl group as defined above where one of more of the hydrogen atoms is replaced by a fluoro. Examples of C

fluoroalkyl include, but are not limited to, -CF3, CF₂H, -C2F5, and -C2F4H. C.3-6 cycloalkyl: the term 'C3-6 cycloalkyl' as used herein, pertains to a monovalent moiety obtained by removing a hydrogen atom from a carbon atom of a saturated cyclic core having 3, 4. 5 or 6 atom in the cyclic core all of which are carbon atoms. Examples of C3-6 cycloalkyl include, but are not limited to, cyclopropyl, cyclohexyl and cyclopentyl.

C5-6 heteroaryl: the term C5-6 heteroaryl as used herein, pertains to a monovalent moiety obtained by removing a hydrogen atom from a ring atom of an aromatic structure having between one and three atoms that are not carbon forming part of said ring. Wherein, those atoms that are not carbon can be chosen independently from the list nitrogen, oxygen and sulphur. The group may be substituted by one or more C a Iky I groups.

Examples of Cs-e heteroaryl groups include, but are not limited to, groups derived from: Ni : pyridine (Ce);

1O1 : oxazole (Cs), isoxazole (Cs);

2O1 : oxadiazole (furazan) (C5);

N2S1 : thiadiazole (C₅)

2: imidazole (1.3-diazole) (C5), pyrazole (1 ,2-diazole) (C5), pyridazine (1 ,2-diazine) (Ce), pyrimidine (1.3-diazine) (Ce) (e.g., cytosine, thymine, uracil), pyrazine (1 ,4-diazine) (Ce); N3: triazole (C5).

C5-6 heteroaryl methyl: -CH2-(Cs-e heteroaryl), wheren Cs-e heteroaryl is as defined above.

C4-6 heterocyclyl: The term "C4-6 heterocyclyl" as used herein, pertains to a monovalent moiety obtained by removing a hydrogen atom from a ring atom of a monocyclic heterocyclic compound, which moiety has from 4 to 6 ring atoms; of which from 1 to 2 atoms are heteroatoms, chosen from oxygen or nitrogen.

In this context, the prefixes (e.g. C₄-e) denote the number of ring atoms, or range of number of ring atoms, whether carbon atoms or heteroatoms.

Examples of C₄-e heterocyclyl groups include, but are not limited to, those derived from: Ni : azetidine (C₄), pyrrolidine (tetrahydropyrrole) (Cs), pyrroline (e.g., 3-pyrroline,

2.5-dihydropyrrole) (Cs), 2H-pyrrole or 3H-pyrrole (isopyrrole, isoazole) (Cs), piperidine (Ce), dihydropyridine (Ce), tetrahydropyridine (Ce), azepine (Or);

2: diazetidine (C₄), imidazolidine (Cs), pyrazolidine (diazolidine) (Cs), imidazoline (Cs), pyrazoline (dihydropyrazole) (Cs), piperazine (Ce); Oi : oxetane (C4), tetrahydrofuran (C5); oxane (Ce);

O2: dioxetane (C4), dioxolane (C5); dioxane (Ce);

1O1 : tetrahydrooxazole (C5), dihydrooxazole (C5), tetrahydroisoxazole (C5),

dihydroisoxazole (Cs), morpholine (Ce), tetrahydrooxazine (Ce), dihydrooxazine (Ce), oxazine (Ce).

Those C4-6 heterocyciyi groups which include a N atom may be substituted on this atom by an acyl group, and in particular, by -C(=0)Me. The C4-6 heterocyciyi groups may also bear a single oxo substituent, as for example in the following group:

C4-6 heterocyciyi methyl: -CH2-(C -e heterocyciyi), wherein C4-6 heterocyciyi is as defined above.

Phenyl: -CeHs, wherein the phenyl may itself be optionally substituted by one or more CM a Iky I groups, one or more C1-4 fluoroalkyl groups, one or more C1-4 alkoxy groups, one or more halo substituents and one or more cyano substituents.

Benzyl: -CH₂-Phenyl, wherein phenyl is as defined above.

Halo: The term "halo" as used herein, refers to a group selected from fluoro, chloro, bromo and iodo.

Amido: -(C=0)NRR' wherein R and R' are independently selected from H, C a Iky I and C4-6 heterocyciyi as defined above, or together form a group selected from (-CH₂-)_n and -(CH₂)_m- X-(CH₂)p-, where n=3-6, m and p=0-4, and X=0 or NH. X may also be N-S(=0)₂, S or S(=0)₂. The cyclic amido groups may also be bridged by a further group selected from (- CH₂-)ni and -(CH₂)_mi-X-(CH₂)_pr, where n1 is 1-3 and ml and p1 are 1 -3. The cyclic amido groups may also be further substituted by one, two or more hydroxy, oxo, Ci-₂ alkyl, Ci.₂ alkoxy, hydroxy-Ci.₂ alkyl, Ci-₂ alkoxy-Ci-₂ alkyl groups or one spiro Ct-e heteroaryl or spiro

and

Amidomethyl: -Ch -amido, where amido is as defined above, Examples of amidomethyl groups include, but are not limited to, -CH₂-C(=0)NH₂, -CH₂-C(=0)NMe₂, -CH₂-

C(=0)NHMe.

Acylamido: -NR(C=0)R^* wherein R and R' are independently selected from H, Ci-t a Iky I and Ci-4 fluoro alkyl as defined above. R' may also be -(CH₂)_n-, where n is 3 or 4. Examples of an acylamido group include, but are not limited to, -N(H)C(=0)CF3, -N(H)C(=0)Me, and:

Acylamidomethyl: -CH₂-acylamido, where acylamido is as defined above, Examples of acylamidomethyl groups include, but are not limited to -CH₂-N(H)C(=0)Me and

-CH₂-N(H)C(=0)CF₃.

Ci-4 a Iky I ester: -C(=0)OR, wherein R is a CM alkyl group. Examples of Ci_i alkyi ester groups include, but are not limited to, -C(=0)OCH₃, -C(=0)OCH₂CH₃, and -C(=0)OC(CH₃)₃. Ci-4 alkyl ester methyl: -CH2-(Ci-4 alkyl ester), where C alkyl ester is as defined above. Examples of C alkyl ester methyl groups include, but are not limited to, -CH₂-C(=0)OCH₃, -CH₂-C(=0)OCH₂CH₃, and -CH₂-C(=0)OC(CH₃)₃.

Ci-i alkyl carbamoyl: -N HC(=0)OR wherein R is a CM alkyl group as defined above.

Examples of C1-4 alkyl carbamoyl include, but are not limited to, -N(H )C(=0)OCH₃,

-N(H )C(=0)OCH₂CH₃, and -N(H )C(=0)OC(CH₃)₃.

C alkyl carbamoyl methyl: -CH₂-(CM alkyl carbamoyl), where CM alkyl carbamoyl is as defined above. Examples of CM alkyl carbamoyl methyl include, but are not limited to, -CH₂-N(H )C(=0)OCH₃, -CH₂-N(H )C(=0)OCH₂CH₃, and -CH₂-N(H )C(=0)OC(CH₃)₃.

C1-4 alkylacyl: -C(=0)R, wherein R is a CM alkyl group as defined above. Examples of CM alkylacyl groups include, but are not limited to, -C(=0)CH₃ (acetyl), -C(=0)CH₂CH₃

(propionyl) and -C(=0)C(CH₃)₃ (t-butyryl).

CM alkylacyl methyl: -CH₂-(CM alkylacyl), where CM alkylacyl is as defined above.

Examples of CM alkylacylmethyl groups include, but are not limited to, -CH₂-C(=0)CH₃, -CH₂-C(=0)CH₂CH₃, and -CH₂-C(=0)C(CH₃)₃. Phenylcarbonyl: -C(=0)-phenyl, where phenyl is as defined above.

Carboxy (carboxylic acid): -C(=0)OH Carboxymethyl: -CH₂-C(=0)OH.

Ether: -OP, wherein P is chosen from one of the following substituents: C1-4 alkyl, benzyl, phenyl, C1-4 fluoroalkyl, Cs-e heteroaryl, -CH2-C5-6 heteroaryl, C4-6 heterocyciyi, and -CH2-C4-6 heterocyciyi as defined above. Examples of an ether include, but are not limited to, -OPh, -OBn, -OCF3, -OCH2CF3, -OCH(CH₃)₂, -OCH₂-cyclopropyl,

-0-(N-acetyl)azetidinyl, e.g. -0-(N-acetyl)piperidinyl, e.g. -O-oxetanyl, e.g.;

Piperidyloxy, e.g -0-(N- O-tetrahydropyranyl, e.g.;

carboxylate)piperidinyl, e.g.

methyl, isopropyl, isobutyl;

-O-tetrahydrofuranyl, e.g.; (8-methoxycarbonyl-8- azabicyclo[3.2.1 ]octan-3- yl)oxy:

Amino: -NPP', wherein P and P' are independently chosen from H, Ci~i alkyl, C4-6 heterocyciyi, phenyl and C5-6 heteroaryl as defined above. Examples of an amine include, but are not limited to, -NH2,-N(H)pyridazinyl,

-NH-(N-acetyl)azetidinyl, e.g. -NH-(N-acetyl)piperidinyl, -NH-oxetanyl, e.g.;

e.g.

(1-methylsulfonylazetidin-3-yl)amino; cyclopentyl(methyl)amino; cyclopentylamino;

Aminomethyl: -Chb-Amino, where amino is as defined above. Examples of aminomethyl include, but are not limited to, -CH2-NH2 and -CH?-N(H)pyridaz!nyl.

Sulfonamido: -SO2N RR' wherein R and R' are independently selected from H, C1-4 alkyl, phenyl and C5-6 heteroaryl as defined above. Examples of sulfonamido groups include, but are not limited to, -S0₂N(Me)₂ and -S0₂NPhMe.

Sulfonamino: -N HSO2R wherein R is selected from C alkyl, phenyl and C5-6 heteroaryl as defined above. Examples of sulfonamino groups include, but are not limited, to -NHSC^Me and -NHS0₂Ph

Sulfone: -SO2R, wherein R is selected from C1-4 alkyl and C1-4 fluoroalkyl as defined above. Example of sulfone groups includes but is not limited to SO2CF3. Sulfoxide: -SOR, wherein R is selected from C1-4 alkyl and C1-4 fluoroalkyl as defined above. Example of sulfoxide groups includes but is not limited to SOCF₃.

Nitrile: -CN. Nitrilemethyl: -CH₂-CN

Fused N-heterocyclic ring: where A is phenyl, it may have a C5-6 Nrcontaining heterocyclic ring fused to it as a substituent group. The C5-6 Nrcontaining heterocyclic ring may in particular be selected from:

Which may be fused in any orientation, and wherein the N ring atom may be optionally substituted, for example by a C1-4 alkylacyl group.

Includes Other Forms

Unless otherwise specified, included in the above are the well known ionic, salt, solvate, and protected forms of these substituents. For example, a reference to carboxylic acid (-COOH) also includes the anionic (carboxylate) form (-COO^"), a salt or solvate thereof, as well as conventional protected forms. Similarly, a reference to an amino group includes the protonated form (-N⁺HR¹ R²), a salt or solvate of the amino group, for example, a hydrochloride salt, as well as conventional protected forms of an amino group. Similarly, a reference to a hydroxyl group also includes the anionic form (-0^"), a salt or solvate thereof, as well as conventional protected forms.

Salts

It may be convenient or desirable to prepare, purify, and/or handle a corresponding salt of the active compound, for example, a pharmaceutically-acceptable salt. Examples of pharmaceutically acceptable salts are discussed in Berge, et al., J. Pharm. Sci., 66, 1 -19 (1977).

For example, if the compound is anionic, or has a functional group which may be anionic (e.g. -COOH may be -COO^"), then a salt may be formed with a suitable cation. Examples of suitable inorganic cations include, but are not limited to, alkali metal ions such as Na ' and K\ alkaline earth cations such as Ca²⁺ and Mg²⁺, and other cations such as ΑΓ³. Examples of suitable organic cations include, but are not limited to, ammonium ion (i.e. NH₄ ⁺) and substituted ammonium ions (e.g. NhfoR^*, H2 2⁺, NHR3', NR. ). Examples of some suitable substituted ammonium ions are those derived from: ethylamine, diethylamine,

dicyclohexylamine, triethylamine, butylamine, ethylenediamine, ethanolamine,

diethanolamine, piperazine, benzylamine, phenylbenzylamine, choline, meglumine, and tromethamine, as well as amino acids, such as lysine and arginine. An example of a common quaternary ammonium ion is N(CH3)₄ ⁺.

If the compound is cationic, or has a functional group which may be cationic (e.g. -NH2 may be -NhV), then a salt may be formed with a suitable anion. Examples of suitable inorganic anions include, but are not limited to, those derived from the following inorganic acids:

hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfurous, nitric, nitrous, phosphoric, and phosphorous. Examples of suitable organic anions include, but are not limited to, those derived from the following organic acids: 2-acetyoxybenzoic, acetic, ascorbic, aspartic, benzoic,

camphorsulfonic, cinnamic, citric, edetic, ethanedisulfonic, ethanesulfonic, fumaric, glucheptonic, gluconic, glutamic, glycolic, hydroxymaleic, hydroxynaphthalene carboxylic, isethionic, lactic, lactobionic, lauric, maleic, malic, methanesulfonic, mucic, oleic, oxalic, palmitic, pamoic, pantothenic, phenylacetic, phenylsulfonic, propionic, pyruvic, salicylic, stearic, succinic, sulfanilic, tartaric, toluenesulfonic, trifluoroacetic acid and valeric. Examples of suitable polymeric organic anions include, but are not limited to, those derived from the following polymeric acids: tannic acid, carboxymethyl cellulose.

Solvates

It may be convenient or desirable to prepare, purify, and/or handle a corresponding solvate of the active compound. The term "solvate" is used herein in the conventional sense to refer to a complex of solute (e.g. active compound, salt of active compound) and solvent. If the solvent is water, the solvate may be conveniently referred to as a hydrate, for example, a mono-hydrate, a di-hydrate, a tri-hydrate, etc.

Isomers

Certain compounds of the invention may exist in one or more particular geometric, optical, enantiomeric, diasteriomeric, epimeric, atropic, stereoisomeric, tautomeric, conformational, or anomeric forms, including but not limited to, cis- and trans-forms; E- and Z-forms; c-, t-, and r- forms; endo- and exo-forms; R-, S-, and meso-forms; D- and L-forms; d- and l-forms; (+) and (-) forms; keto-, enol-, and enolate-forms; syn- and anti-forms; synclinal- and anticlinal-forms; a- and β-forms; axial and equatorial forms; boat-, chair-, twist-, envelope-, and halfchair-forms; and combinations thereof, hereinafter collectively referred to as

"isomers" (or "isomeric forms").

The term "chirai" refers to molecules which have the property of non-superimposability of the mirror image partner, while the term "achiral" refers to molecules which are superimposable on their mirror image partner. The term "stereoisomers" refers to compounds which have identical chemical constitution, but differ with regard to the arrangement of the atoms or groups in space.

"Diastereomer" refers to a stereoisomer with two or more centers of chirality and whose molecules are not mirror images of one another. Diastereomers have different physical properties, e.g. melting points, boiling points, spectral properties, and reactivities. Mixtures of diastereomers may separate under high resolution analytical procedures such as electrophoresis and chromatography.

"Enantiomers" refer to two stereoisomers of a compound which are non-superimposable mirror images of one another. Stereochemical definitions and conventions used herein generally follow S. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994. The compounds of the invention may contain asymmetric or chiral centers, and therefore exist in different stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds of the invention, including but not limited to, diastereomers, enantiomers and atropisomers, as well as mixtures thereof such as racemic mixtures, form part of the present invention. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. In describing an optically active compound, the prefixes D and L, or R and S, are used to denote the absolute configuration of the molecule about its chiral center(s). The prefixes d and I or (+) and (-) are employed to designate the sign of rotation of plane-polarized light by the compound, with (-) or I meaning that the compound is levorotatory. A compound prefixed with (+) or d is dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of one another. A specific stereoisomer may also be referred to as an enantiomer, and a mixture of such isomers is often called an enantiomeric mixture. A 50:50 mixture of enantiomers is referred to as a racemic mixture or a racemate, which may occur where there has been no stereoselection or stereospecificity in a chemical reaction or process. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomeric species, devoid of optical activity.

Note that, except as discussed below for tautomeric forms, specifically excluded from the term "isomers", as used herein, are structural (or constitutional) isomers (i.e. isomers which differ in the connections between atoms rather than merely by the position of atoms in space). For example, a reference to a methoxy group, -OCH3, is not to be construed as a reference to its structural isomer, a hydroxymethyl group, -CH2OH. Similarly, a reference to ortho-chlorophenyl is not to be construed as a reference to its structural isomer, meta- chlorophenyl. However, a reference to a class of structures may well include structurally isomeric forms falling within that class (e.g. C1-7 alkyl includes n-propyl and iso-propyl; butyl includes n-, iso-, sec-, and tert-butyl; methoxyphenyl includes ortho-, meta-, and para- methoxyphenyl).

The above exclusion does not pertain to tautomeric forms, for example, keto-, enol-, and enolate-forms, as in, for example, the following tautomeric pairs: keto/enol (illustrated below), imine/enamine, amide/imino alcohol, amidine/amidine, nitroso/oxime,

thioketone/enethiol, N-nitroso/hyroxyazo, and nitro/aci-nitro.

keto enol enolate

The term "tautomer" or "tautomeric form" refers to structural isomers of different energies which are interconvertible via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via migration of a proton, such as keto-enol and imine-enamine isomerizations. Valence tautomers include interconversions by reorganization of some of the bonding electrons.

Note that specifically included in the term "isomer" are compounds with one or more isotopic substitutions. For example, H may be in any isotopic form, including ¹ H, ²H (D), and ³H (T); C may be in any isotopic form, including ¹²C, ¹³C, and ¹⁴C; O may be in any isotopic form, including ¹⁶0 and ¹⁸0; and the like.

Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, and chlorine, such as, but not limited to ²H (deuterium, D), ³H (tritium), C, ³C, ¹⁴C, ¹⁵N, ¹⁸F, ³ P, ³²P, ³⁵S, ³⁶CI, and ^{1 5}l. Various isotopically labeled compounds of the present invention, for example those into which radioactive isotopes such as 3H, 13C, and 14C are incorporated. Such isotopically labelled compounds may be useful in metabolic studies, reaction kinetic studies, detection or imaging techniques, such as positron emission tomography (PET) or single- photon emission computed tomography (SPECT) including drug or substrate tissue distribution assays, or in radioactive treatment of patients. Deuterium labelled or substituted therapeutic compounds of the invention may have improved DMPK (drug metabolism and pharmacokinetics) properties, relating to distribution, metabolism, and excretion (ADME). Substitution with heavier isotopes such as deuterium may afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements. An 18F labeled compound may be useful for PET or SPECT studies. Isotopically labeled compounds of this invention and prodrugs thereof can generally be prepared by carrying out the procedures disclosed in the schemes or in the examples and preparations described below by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent. Further, substitution with heavier isotopes, particularly deuterium (i.e., 2H or D) may afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements or an improvement in therapeutic index. It is understood that deuterium in this context is regarded as a substituent. The concentration of such a heavier isotope, specifically deuterium, may be defined by an isotopic enrichment factor. In the compounds of this invention any atom not specifically designated as a particular isotope is meant to represent any stable isotope of that atom.

Unless otherwise specified, a reference to a particular compound includes all such isomeric forms, including (wholly or partially) racemic and other mixtures thereof. Methods for the preparation (e.g. asymmetric synthesis) and separation (e.g. fractional crystallisation and chromatographic means) of such isomeric forms are either known in the art or are readily obtained by adapting the methods taught herein, or known methods, in a known manner.

Therapeutic Indications

Compounds disclosed herein may provide a therapeutic benefit in a number of disorders, in particular, in the treatment or prevention of cancers and hemoglobinopathies.

Cancer

Modulators of PRMT5 mediated post-translational arginine methylation of p53 may regulate a pro-apoptotic p53 response, and may therefore be useful as therapeutic agents, for example in the treatment of cancer. Such agents may also be useful as therapeutic agents for the treatment of cancers which exhibit overexpression of PRMT5.

A "cancer" may be any form of cancer. In particular, a cancer can comprise any one or more of the following: leukemia, acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), non-Hodgkin's lymphoma, Hodgkin's disease, prostate cancer, lung cancer, melanoma, breast cancer, colon and rectal cancer, colon cancer, squamous cell carcinoma and gastric cancer.

Alternatively, the cancer may comprise adrenocortical cancer, anal cancer, bladder cancer, blood cancer, bone cancer, brain tumor, cancer of the female genital system, cancer of the male genital system, central nervous system lymphoma, cervical cancer, childhood rhabdomyosarcoma, childhood sarcoma, endometrial cancer, endometrial sarcoma, esophageal cancer, eye cancer, gallbladder cancer, gastrointestinal tract cancer, hairy cell leukemia, head and neck cancer, hepatocellular cancer, hypopharyngeal cancer, Kaposi's sarcoma, kidney cancer, laryngeal cancer, liver cancer, malignant fibrous histiocytoma, malignant thymoma, mesothelioma, multiple myeloma, myeloma, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, nervous system cancer, neuroblastoma, oral cavity cancer, oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pituitary tumor, plasma cell neoplasm, primary CNS lymphoma, rectal cancer, respiratory system, retinoblastoma, salivary gland cancer, skin cancer, small intestine cancer, soft tissue sarcoma, stomach cancer, stomach cancer, testicular cancer, thyroid cancer, urinary system cancer, uterine sarcoma, vaginal cancer, vascular system, Waldenstrom's macroglobulinemia and/or Wilms' tumor.

Cancers may be of a particular type. Examples of types of cancer include lymphoma, melanoma, carcinoma (e.g. adenocarcinoma, hepatocellular carcinoma, medullary carcinoma, papillary carcinoma, squamous cell carcinoma), astrocytoma, glioma, medulloblastoma, myeloma, meningioma, neuroblastoma, sarcoma (e.g. angiosarcoma, chrondrosarcoma, osteosarcoma).

The cancer may be a PRMT5 overexpressing cancer. The cancer may over express PRMT5 protein relative to non-cancerous tissue. In some cases, the cancer overproduces PRMT5 mRNA relative to non-cancerous tissue.

Alternatively or additionally, the cancer may be a p53 overexpressing cancer. The cell may overexpress p53 protein relative to non-cancerous tissue. It may overproduce p53 mRNA as compared to non-cancerous tissue. In some cases, the level of p53 protein and/or mRNA in the cell is at a level approximately equivalent to that of a non-cancerous cell.

The agents described herein may be useful in combination with other anti-cancer therapies. They may act synergistically with chemo- or radiotherapy, and/or with p53 targeted drugs.

An inhibitor of PRMT5 would in all likelihood augment the effects of drugs (such as the nutlins) that restore p53. Inhibition of PRMT5, resulting in decreased arginine-methylated p53, may sensitize tumour cells to chemo- and radiotherapy by switching, or at least biasing, the cellular outcome to apoptosis.

Combination therapies

p53 is activated by DNA damage. PRMT5 is part of the complex of proteins that activate and modulate p53 activity in response to DNA damage. It is likely that inhibition of PRMT5, resulting in decreased arginine-methylated p53, would sensitize tumour cells to chemo- and radiotherapy by switching or at least biasing the cellular outcome to apoptosis. PRMT5 inhibition is likely to synergize well with low dose chemo- or radiotherapy, by stabilizing p53, and biasing the cellular outcome to apoptosis.

Biasing the p53 response towards apoptosis would in all likelihood be of benefit, and an agent that so biases the response would be expected to augment the effect of a p53 resurrecting drug. Thus, in some cases, a PRMT5 modulator disclosed herein may be administered in conjunction with a radiotherapeutic or chemotherapeutic regime. It may be administered in conjunction with a drug that resurrects cellular p53 activity, for example, a p53 agonist. The PRMT5 modulator may be administered simultaneously or sequentially with radio and/or chemotherapy. Suitable chemotherapeutic agents and radiotherapy protocols will be readily appreciable to the skilled person. In particular, the compound described herein may be combined with low dose chemo or radio therapy. Appropriate dosages for low dose" chemo or radio therapy will be readily appreciable to the skilled practitioner.

Hemoglobinopathies

The compounds disclosed herein may be useful in the treatment or prevention of conditions that may benefit from the increased expression of γ-globin genes, for example, due to the release of repressive methylation of these genes. The compounds disclosed herein may be useful in the treatment or prevention of hemoglobinopathies. A hemoglobinopathy is a condition associated with the presence of abnormal hemoglobin in the blood of a subject. Such conditions include β-thalassemia and Sickle Cell Disease, a-thalassemia and δ- thalassemia. Hemoglobinopathies treatable by the compounds disclosed herein may be ameliorated by the re-activation of the subjects γ-globin genes (γ genes). In such cases, the subject is not a fetal mammal.

Methods of Treatment

The compounds of the present invention may be used in a method of therapy. Also provided is a method of treatment, comprising administering to a subject in need of treatment a therapeutically-effective amount of a compound of the invention. The term "therapeutically effective amount" is an amount sufficient to show benefit to a patient. Such benefit may be at least amelioration of at least one symptom. The actual amount administered, and rate and time-course of administration, will depend on the nature and severity of what is being treated. Prescription of treatment, e.g. decisions on dosage, is within the responsibility of general practitioners and other medical doctors.

As described above, the anti cancer treatment defined herein may be applied as a sole therapy or may involve, in addition to the compound of the invention, conventional surgery or radiotherapy or chemotherapy. Such chemotherapy may include one or more of the following categories of anti-tumour agents:-

(i) other antiproliferative/antineoplastic drugs and combinations thereof, as used in medical oncology, such as alkylating agents (for example cisplatin, oxaliplatin, carboplatin, cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulphan, temozolamide and nitrosoureas); antimetabolites (for example gemcitabine and antifolates such as fluoropyrimidines like 5 fluorouracil and tegafur, raltitrexed, methotrexate, cytosine arabinoside, and hydroxyurea); antitumour antibiotics (for example anthracyclines like adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin and mithramycin); antimitotic agents (for example vinca alkaloids like vincristine, vinblastine, vindesine and vinorelbine and taxoids like taxol and docetaxel (Taxotere) and polokinase inhibitors); and topoisomerase inhibitors (for example

epipodophyllotoxins like etoposide and teniposide, amsacrine, topotecan and camptothecin);

(ii) cytostatic agents such as antioestrogens (for example tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene and iodoxyfene), antiandrogens (for example

bicalutamide, flutamide, nilutamide and cyproterone acetate), LHRH antagonists or LHRH agonists (for example goserelin, leuprorelin and buserelin), progestogens (for example megestrol acetate), aromatase inhibitors (for example as anastrozole, letrozole, vorazole and exemestane) and inhibitors of 5^*-reductase such as finasteride;

(iii) anti-invasion agents (for example c-Src kinase family inhibitors like 4-(6-chloro-2.3- methylenedioxyanilino)-7-[2-(4-methylpiperazin-1 -yl)ethoxy]-5-tetrahydropyran-4- yloxyquinazoline (AZD0530; International Patent Application WO 01/94341 ), N-(2-chloro-6- methylphenyl)-2-{6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-methylpyrimidin-4-ylamino}thiazole- 5-carboxamide (dasatinib, BMS-354825; J. Med. Chem., 2004, 47, 6658-6661 and and 4- ((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxy-7-(3-(4-methylpiperazin-1 - yl)propoxy)quinoline-3-carbonitrile (bosutinib, SKI-606; Cancer research (2003), 63(2), 375- 81 ), and metalloproteinase inhibitors like marimastat, inhibitors of urokinase plasminogen activator receptor function or antibodies to Heparanase);

(iv) inhibitors of growth factor function: for example such inhibitors include growth factor antibodies and growth factor receptor antibodies (for example the anti erbB2 antibody trastuzumab [HerceptinT], the anti-EGFR antibody panitumumab, the anti erbB1 antibody cetuximab [Erbitux, C225] and any growth factor or growth factor receptor antibodies disclosed by Stern et al. Critical reviews in oncology/haematology, 2005, Vol. 54, pp1 1-29); such inhibitors also include tyrosine kinase inhibitors, for example inhibitors of the epidermal growth factor family (for example EGFR family tyrosine kinase inhibitors such as

N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4-amine

(gefitinib, ZD1839), N-(3-ethynylphenyl)-6.7-bis(2-methoxyethoxy)quinazolin-4-amine (erlotinib, OSI 774) and 6-acrylamido-N-(3-chloro-4-fiuorophenyl)-7-(3-morpholinopropoxy)- quinazolin-4-amine (CI 1033), erbB2 tyrosine kinase inhibitors such as lapatinib, inhibitors of the hepatocyte growth factor family, inhibitors of the platelet-derived growth factor family such as imatinib, inhibitors of serine/threonine kinases (for example Ras/Raf signalling inhibitors such as farnesyl transferase inhibitors, for example sorafenib (BAY 43-9006)), inhibitors of cell signalling through MEK and/or AKT kinases, inhibitors of the hepatocyte growth factor family, c-kit inhibitors, abl kinase inhibitors, IGF receptor (insulin-like growth factor) kinase inhibitors; aurora kinase inhibitors (for example AZD1 152, PH739358, VX-680, MLN8054, R763, MP235, MP529, VX-528 AND AX39459) and cyclin dependent kinase inhibitors such as CDK2 and/or CDK4 inhibitors;

(v) antiangiogenic and antilymphangiogenic agents such as those which inhibit the effects of vascular endothelial growth factor, [for example the anti vascular endothelial cell growth factor A (VEGFA) antibody bevacizumab (AvastinT), the anti vascular endothelial cell growth factor A (VEGFA) antibody ranibizumab, the anti-VEGF aptamer pegaptanib, the anti vascular endothelial growth factor receptor 3 (VEGFR3) antibody IMC-3C5, the anti vascular endothelial cell growth factor C (VEGFC) antibody VGX-100, the anti vascular endothelial cell growth factor D (VEGFD) antibody VGX-200, the soluble form of the vascular endothelial growth factor receptor 3 (VEGFR3) VGX-300 and VEGF receptor tyrosine kinase inhibitors such as 4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline (vandetanib; ZD6474; Example 2 within WO 01 /32651 ), 4-(4-fluoro-2-methylindol-5-yloxy)-6- methoxy-7-(3-pyrrolidin-1 -ylpropoxy)quinazoline (cediranib; AZD2171 ; Example 240 within WO 00/47212), vatalanib (PTK787; WO 98/35985), pazopanib (GW786034), axitinib

(AG013736), sorafenib and sunitinib (SU1 1248; WO 01/60814), compounds such as those disclosed in International Patent Applications W097/22596, WO 97/30035, WO 97/32856 and WO 98/13354 and compounds that work by other mechanisms (for example linomide, inhibitors of integrin avb3 function and angiostatin)];

(vi) vascular damaging agents such as Combretastatin A4 and compounds disclosed in International Patent Applications WO 99/02166, WO 00/40529, WO 00/41669, WO

01/92224, WO 02/04434 and WO 02/08213; (vii) antisense therapies, for example those which are directed to the targets listed above, such as ISIS 2503, an anti-ras antisense;

(viii) gene therapy approaches, including for example approaches to replace aberrant genes such as aberrant p53 or aberrant BRCA1 or BRCA2, GDEPT (gene directed enzyme pro drug therapy) approaches such as those using cytosine deaminase, thymidine kinase or a bacterial nitroreductase enzyme and approaches to increase patient tolerance to chemotherapy or radiotherapy such as multi drug resistance gene therapy; and

(ix) immunotherapy approaches, including for example ex vivo and in vivo approaches to increase the immunogenicity of patient tumour cells, such as transfection with cytokines such as interleukin 2, interleukin 4 or granulocyte macrophage colony stimulating factor, approaches to decrease T cell anergy, approaches using transfected immune cells such as cytokine transfected dendritic cells, approaches using cytokine transfected tumour cell lines and approaches using anti idiotypic antibodies Administration

The active compound or pharmaceutical composition comprising the active compound may be administered to a subject by any convenient route of administration, whether

systemically/ peripherally or at the site of desired action, including but not limited to, oral (e.g. by ingestion); topical (including e.g. transdermal, intranasal, ocular, buccal, and sublingual); pulmonary (e.g. by inhalation or insufflation therapy using, e.g. an aerosol, e.g. through mouth or nose); rectal; vaginal; parenteral, for example, by injection, including subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratracheal,

subcuticular, intraarticular, subarachnoid, intravitreal and intrasternal; by implant of a depot, for example, subcutaneously, intravitreal or intramuscularly. The subject may be a eukaryote, an animal, a vertebrate animal, a mammal, a rodent (e.g. a guinea pig, a hamster, a rat, a mouse), murine (e.g. a mouse), canine (e.g. a dog), feline (e.g. a cat), equine (e.g. a horse), a primate, simian (e.g. a monkey or ape), a monkey (e.g. marmoset, baboon), an ape (e.g. gorilla, chimpanzee, orang-utan, gibbon), or a human.

Formulations

While it is possible for the active compound to be administered alone, it is preferable to present it as a pharmaceutical composition (e.g. formulation) comprising at least one active compound, as defined above, together with one or more pharmaceutically acceptable carriers, adjuvants, excipients, diluents, fillers, buffers, stabilisers, preservatives, lubricants, or other materials well known to those skilled in the art and optionally other therapeutic or prophylactic agents.

Thus, the present invention further provides pharmaceutical compositions, as defined above, and methods of making a pharmaceutical composition comprising admixing at least one active compound, as defined above, together with one or more pharmaceutically acceptable carriers, excipients, buffers, adjuvants, stabilisers, or other materials, as described herein.

The term "pharmaceutically acceptable" as used herein pertains to compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgement, suitable for use in contact with the tissues of a subject (e.g. human) without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio. Each carrier, excipient, etc. must also be "acceptable" in the sense of being compatible with the other ingredients of the formulation.

Suitable carriers, excipients, etc. can be found in standard pharmaceutical texts, for example, Remington's Pharmaceutical Sciences, 18th edition, Mack Publishing Company, Easton, Pa., 1990.

The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. Such methods include the step of bringing into association the active compound with the carrier which constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the active compound with liquid carriers or finely divided solid carriers or both, and then if necessary shaping the product.

Formulations may be in the form of liquids, solutions, suspensions, emulsions, elixirs, syrups, tablets, losenges, granules, powders, capsules, cachets, pills, ampoules, suppositories, pessaries, ointments, gels, pastes, creams, sprays, mists, foams, lotions, oils, boluses, electuaries, or aerosols.

Formulations suitable for oral administration (e.g. by ingestion) may be presented as discrete units such as capsules, cachets or tablets, each containing a predetermined amount of the active compound; as a powder or granules; as a solution or suspension in an aqueous or non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion; as a bolus; as an electuary; or as a paste. A tablet may be made by conventional means, e.g., compression or moulding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active compound in a free-flowing form such as a powder or granules, optionally mixed with one or more binders (e.g. povidone, gelatin, acacia, sorbitol, tragacanth, hydroxypropylmethyl cellulose); fillers or diluents (e.g. lactose, microcrystalline cellulose, calcium hydrogen phosphate); lubricants (e.g. magnesium stearate, talc, silica); disintegrants (e.g. sodium starch glycolate, cross-linked povidone, cross-linked sodium carboxymethyl cellulose); surface-active or dispersing or wetting agents (e.g. sodium lauryl sulfate); and preservatives (e.g. methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, sorbic acid). Moulded tablets may be made by moulding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be coated or scored and may be formulated so as to provide slow or controlled release of the active compound therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile. Tablets may optionally be provided with an enteric coating, to provide release in parts of the gut other than the stomach.

Formulations suitable for topical administration (e.g. transdermal, intranasal, ocular, buccal, and sublingual) may be formulated as an ointment, cream, suspension, lotion, powder, solution, past, gel, spray, aerosol, or oil. Alternatively, a formulation may comprise a patch or a dressing such as a bandage or adhesive plaster impregnated with active compounds and optionally one or more excipients or diluents.

Formulations suitable for topical administration in the mouth include losenges comprising the active compound in a flavoured basis, usually sucrose and acacia or tragacanth; pastilles comprising the active compound in an inert basis such as gelatin and glycerin, or sucrose and acacia; and mouthwashes comprising the active compound in a suitable liquid carrier.

Formulations suitable for topical administration to the eye also include eye drops wherein the active compound is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the active compound.

Formulations suitable for nasal administration, wherein the carrier is a solid, include a coarse powder having a particle size, for example, in the range of about 20 to about 500 microns which is administered in the manner in which snuff is taken, i.e. by rapid inhalation through the nasal passage from a container of the powder held close up to the nose. Suitable formulations wherein the carrier is a liquid for administration as, for example, nasal spray, nasal drops, or by aerosol administration by nebuliser, include aqueous or oily solutions of the active compound. Formulations suitable for administration by inhalation include those presented as an aerosol spray from a pressurised pack, with the use of a suitable propellant, such as

dichlorodifluoromethane, trichlorofluoromethane, dichoro-tetrafluoroethane, carbon dioxide, or other suitable gases.

Formulations suitable for topical administration via the skin include ointments, creams, and emulsions. When formulated in an ointment, the active compound may optionally be employed with either a paraffinic or a water-miscible ointment base. Alternatively, the active compounds may be formulated in a cream with an oil-in-water cream base. If desired, the aqueous phase of the cream base may include, for example, at least about 30% w/w of a polyhydric alcohol, i.e., an alcohol having two or more hydroxyl groups such as propylene glycol, butane-1.3-diol, mannitol, sorbitol, glycerol and polyethylene glycol and mixtures thereof. The topical formulations may desirably include a compound which enhances absorption or penetration of the active compound through the skin or other affected areas. Examples of such dermal penetration enhancers include dimethylsulfoxide and related analogues.

When formulated as a topical emulsion, the oily phase may optionally comprise merely an emulsifier (otherwise known as an emulgent), or it may comprises a mixture of at least one emulsifier with a fat or an oil or with both a fat and an oil. Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier which acts as a stabiliser. It is also preferred to include both an oil and a fat. Together, the emulsifier(s) with or without stabiliser(s) make up the so-called emulsifying wax, and the wax together with the oil and/or fat make up the so-called emulsifying ointment base which forms the oily dispersed phase of the cream formulations.

Suitable emulgents and emulsion stabilisers include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl monostearate and sodium lauryl sulphate. The choice of suitable oils or fats for the formulation is based on achieving the desired cosmetic properties, since the solubility of the active compound in most oils likely to be used in pharmaceutical emulsion formulations may be very low. Thus the cream should preferably be a non-greasy, non-staining and washable product with suitable consistency to avoid leakage from tubes or other containers. Straight or branched chain, mono- or dibasic alkyl esters such as di- isoadipate, isocetyl stearate, propylene glycol diester of coconut fatty acids, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate or a blend of branched chain esters known as Crodamol CAP may be used, the last three being preferred esters. These may be used alone or in combination depending on the properties required.

Alternatively, high melting point lipids such as white soft paraffin and/or liquid paraffin or other mineral oils can be used.

Formulations suitable for rectal administration may be presented as a suppository with a suitable base comprising, for example, cocoa butter or a salicylate.

Formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or spray formulations containing in addition to the active compound, such carriers as are known in the art to be appropriate.

Formulations suitable for parenteral administration (e.g. by injection, including cutaneous, subcutaneous, intramuscular, intravenous and intradermal), include aqueous and non- aqueous isotonic, pyrogen-free, sterile injection solutions which may contain anti-oxidants, buffers, preservatives, stabilisers, bacteriostats, and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents, and liposomes or other microparticulate systems which are designed to target the compound to blood components or one or more organs. Examples of suitable isotonic vehicles for use in such formulations include Sodium Chloride Injection, Ringer's Solution, or Lactated Ringer's Injection. Typically, the concentration of the active compound in the solution is from about 1 ng/mL to about 10 g/mL, for example from about 10 ng/ml to about 1 pg/mL The formulations may be presented in unit-dose or multi-dose sealed containers, for example, ampoules and vials, and may be stored in a freeze-dried (lyophilised) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets. Formulations may be in the form of liposomes or other microparticulate systems which are designed to target the active compound to blood components or one or more organs. Dosage

It will be appreciated by one of skill in the art that appropriate dosages of the compound, and compositions comprising the compound, can vary from patient to patient. Determining the optimal dosage will generally involve the balancing of the level of therapeutic benefit against any risk or deleterious side effects. The selected dosage level will depend on a variety of factors including, but not limited to, the activity of the particular compound, the route of administration, the time of administration, the rate of excretion of the compound, the duration of the treatment, other drugs, compounds, and/or materials used in combination, the severity of the condition, and the species, sex, age, weight, condition, general health, and prior medical history of the patient. The amount of compound and route of administration will ultimately be at the discretion of the physician, veterinarian, or clinician, although generally the dosage will be selected to achieve local concentrations at the site of action which achieve the desired effect without causing substantial harmful or deleterious side-effects. Administration can be effected in one dose, continuously or intermittently (e.g., in divided doses at appropriate intervals) throughout the course of treatment. Methods of determining the most effective means and dosage of administration are well known to those of skill in the art and will vary with the formulation used for therapy, the purpose of the therapy, the target cell(s) being treated, and the subject being treated. Single or multiple administrations can be carried out with the dose level and pattern being selected by the treating physician, veterinarian, or clinician.

In general, a suitable dose of the active compound is in the range of about 100 ng to about 25 mg (more typically about 1 pg to about 10 mg) per kilogram body weight of the subject per day. Where the active compound is a salt, an ester, an amide, a prodrug, or the like, the amount administered is calculated on the basis of the parent compound and so the actual weight to be used is increased proportionately.

In one embodiment, the active compound is administered to a human patient according to the following dosage regime: about 100 mg, 3 times daily.

In one embodiment, the active compound is administered to a human patient according to the following dosage regime: about 150 mg, 2 times daily. In one embodiment, the active compound is administered to a human patient according to the following dosage regime: about 200 mg, 2 times daily. However in one embodiment, the active compound is administered to a human patient according to the following dosage regime: about 50 or about 75 mg, 3 or 4 times daily. In one embodiment, the active compound is administered to a human patient according to the following dosage regime: about 100 or about 125 mg, 2 times daily.

Treatment

The term "treatment," as used herein in the context of treating a condition, pertains generally to treatment and therapy, whether of a human or an animal (e.g., in veterinary applications), in which some desired therapeutic effect is achieved, for example, the inhibition of the progress of the condition, and includes a reduction in the rate of progress, a halt in the rate of progress, regression of the condition, amelioration of the condition, and cure of the condition. Treatment as a prophylactic measure (i.e., prophylaxis, prevention) is also included.

The term "therapeutically-effective amount," as used herein, pertains to that amount of an active compound, or a material, composition or dosage from comprising an active

compound, which is effective for producing some desired therapeutic effect, commensurate with a reasonable benefit/risk ratio, when administered in accordance with a desired treatment regimen.

Similarly, the term "prophylactically-effective amount," as used herein, pertains to that amount of an active compound, or a material, composition or dosage from comprising an active compound, which is effective for producing some desired prophylactic effect, commensurate with a reasonable benefit/risk ratio, when administered in accordance with a desired treatment regimen.

The Subject/Patient

The subject/patient may be an animal, mammal, a placental mammal, a marsupial

(e.g., kangaroo, wombat), a monotreme (e.g., duckbilled platypus), a rodent (e.g., a guinea pig, a hamster, a rat, a mouse), murine (e.g., a mouse), a lagomorph (e.g., a rabbit), avian (e.g., a bird), canine (e.g., a dog), feline (e.g., a cat), equine (e.g., a horse), porcine (e.g., a pig), ovine (e.g., a sheep), bovine (e.g., a cow), a primate, simian (e.g., a monkey or ape), a monkey (e.g., marmoset, baboon), an ape (e.g., gorilla, chimpanzee, orangutang, gibbon), or a human. Furthermore, the subject/patient may be any of its forms of development, for example, a foetus. In one preferred embodiment, the subject/patient is a human. General synthesis methods

The compounds of the invention can be prepared employing the following general methods a nd using procedures described in detail in the examples. The reaction conditions referred to are illustrative and non-limiting, for example one skilled in the art may use a diverse range of synthetic methods to synthesis the desired compounds such as but not limited to methods described in literature (for example but not limited to March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 7th Edition or Larock's Comprehensive Organic Transformations: Comprehensive Organic Transformations: A Guide to Functional Group Preparations). Compounds of formula I (la, lb and lc), as described above, can be prepared by synthetic strategies outlined below, wherein the definitions above apply. The synthetic strategies could be applied to the use of racemic or single enantiomer starting materials.

General synthesis method 1

Scheme 1 A illustrates the formation of the amide bond by coupling a relevant carboxylic acid to a primary amine or a secondary amine G1. Methods to form such amides will be apparent to those skilled in the art, but include for example the use of reagents such as HATU, HBTU, T3P and EDCI/HOBt, and the use of activated forms of the carboxylic acid such as the corresponding acyl halide, mixed anhydride or A/-hydroxysuccinimide ester.

G1 G2

Scheme 1A

R⁷ represents the fused cyclic system.

Optionally, where R^N = H, a protecting group (PG) may be used such as but not limited to Boc, allyl, benzyl, 2.4-dimethoxybenzyl. As illustrated in Scheme 1 B, coupling between a relevant carboxylic acid or activated acid, e.g. acyl halide to a primary amine or a secondary amine G3 bearing a protecting group (PG) is performed as described previously. Conditions for the removal of the protecting group are dependent on the type of protecting group employed, and may include but is not limited to such methods as acid hydrolysis, transition metal catalysed cleavage and hydrogenation over transition metal catalysts. Other suitable protecting groups and removal methods will be known to those skilled in the art (for example Greene's Protective Groups in Organic Synthesis, 4th Edition). The use of such a protecting group could be relevant in the other Schemes described.

G4 G2'

Scheme 1 B

Where A contains a direct carboxylic acid substitution or a further substitution that also has a carboxylic acid substitution G5, another amide formation can be conducted to provide compounds of G6, scheme 1 C. Coupling is possible with a primary or secondary amine or cyclised secondary amine including, but not limited to, azetidine, pyrrolidine, piperidine, piperazine and morpholine or substituted analgoues thereof.

Scheme 1C

General synthesis method 2

Scheme 2A illustrates the synthesis of the substituted amine alcohol G9. This is achieved by opening the epoxide with a desired amine (HNR^NR⁷) to form the intermediate G7. The phthalimide protecting group can then be removed by heating with hydrazine hydrate to form G8. Other suitable protecting groups and removal methods will be known to those skilled in the art (for example Greene's Protective Groups in Organic Synthesis, 4th Edition).

Commercially available enantiomerically pure epoxides could be used to give single enantiomer products G9.

Scheme 2A

The amide formation to form G9 can be achieved by the methods outline in Scheme 1A. The synthesis of either enantiomer and the racemate can be achieved by the same method. General synthesis method 3

Scheme 3A

Where R^7a and R^7b together with the atom to which they are bound form R⁷. Scheme 3A illustrates the synthesis of compounds with the formula G13, beginning with an amide bond formation by coupling a relevant carboxylic acid to a primary amine or a secondary amine G10. Methods to form such amides will be apparent to those skilled in the art, but include for example the use of reagents such as HATU, HBTU, T3P and EDCI/HOBt, and the use of activated forms of the carboxylic acid such as the corresponding acyl halide, mixed anhydride or V-hydroxysuccinimide ester.

Suitably protected amino groups and methods for the removal of said protecting groups will be known to those skilled in the art (for example Greene's Protective Groups in Organic Synthesis, 4th Edition). Suitable protecting groups might include Boc, CBz, or phthalimide. Upon removal of the protecting group to provide compounds of the general formula G12, these intermediates can be converted to the desired compound, G13 Methods to form such an intermediate will be apparent to those skilled in the art, but include for example reductive amination by the use of reagents such as, but not limited to sodium triacetoxyborohydride or sodium cyanoborohydride with acetic acid, and sodium triacetoxyborohydride

triacetoxyborohydride or sodium borohydride with Ti(0'Pr)4.

General synthesis method 4

G15 G16

Scheme 4A

Scheme 4A illustrates an approach to the synthesis of compounds with the formula of G16, beginning with the reaction of the desired amine (HNR^NR⁷) with an epoxide bearing a suitable protected amino group such as for example iert-butyl A/-(oxiran-2-yi methyl) carbamate. Opening of the epoxide under suitable conditions furnishes the intermediate compound G14. Suitably protected amino groups and methods for the removal of said protecting groups will be known to those skilled in the art (for example Greene's Protective Groups in Organic Synthesis, 4th Edition). Upon removal of the protecting group to provide compounds of the general formula G15, these intermediates can be converted to the desired compound, G16, by the procedure outlined in Scheme 1A.

General synthesis method 5

Scheme 5A illustrates how to form amine substitutions such as shown in G17. An amide is reacted with a halo-epoxide or similar and the resultant mixture reacted on with a desired amine (HNR⁶R⁷) resulting in a product within the scope of G17. The group denoted (X) can be but is not limited to halogen, tosylate, nosylate or similar.

G17

Scheme 5A

Alternatively enantiomerically pure forms of for example epichlorohydrin can be used to obtain the products G17 in enantiomerically pure form.

General synthesis method 6

Scheme 6A illustrates how to form amine substitutions such as shown in G19. Suitable amide protecting groups and methods for the removal of said protecting groups will be known to those skilled in the art (for example Greene's Protective Groups in Organic

Synthesis, 4th Edition). An amide is reacted with a halo-epoxide or similar and the resultant mixture reacted on with a desired amine (HNR^NR⁷) resulting in an intermediate G18. The group denoted (X) can be but is not limited to halogen, tosylate, nosylate or similar. Removal of the protecting group provides compounds of the general formula G19

G18 G19

Scheme 6A

Alternatively enantiomerically pure forms of the epoxide for example include but are not limited to epichlorohydrin, can be used to obtain the products G14 in enantiomerically pure form.

General synthesis method 7

Scheme 7 A illustrates an approach to the synthesis of compounds with the formula of G23, beginning with the reaction of the desired amine (HNR^NR⁷) with an epoxide bearing a leaving group (LG). The group represented by (LG) includes but not limited to halide, mesylate, tosylate, nosylate. Depending on the conditions and the nature of the leaving group, the reaction can proceed through two different pathways to give either epoxide with structure G20 or alcohol with structure G21. Treatment of either compound with primary amine (R⁵- NH2) furnishes intermediates with structure G22. These intermediates can be converted to the desired com ound, G23, by the procedure outlined in Scheme 1 A.

G23

Scheme 7 A General synthesis method 8

Scheme 8A illustrates the synthesis of compounds G27 which begins with a coupling of a compound G24 with a desired amine (HNR^NR⁷) by substitution of a leaving group (LG) to give intermediates G25. The group represented by (LG) includes but is not limited to halide, mesylate, tosylate, nosylate. The group represented by (PG) is a suitable amine protecting group which includes but is not limited to Boc, phthalimide, benzyl, PMB, allyl. Suitable amine protecting groups and methods for the removal of said protecting groups will be known to those skilled in the art (for example Greene's Protective Groups in Organic Synthesis, 4th Edition). Upon removal of the protecting group, coupling with a suitable carboxylic acid can be performed by methods illustrated in Scheme 1 A.

G26 G27

Scheme 8A

Alternatively, as shown in Scheme 8B, compounds with structure G28 can be coupled with the desired amine (HNR^NR⁷) to give compounds G27.

G28 G27

Scheme 8B

General synthesis method 9

Scheme 9A illustrates the synthesis of compounds G32 where R^3a and R^3b are hydrogen. Amide bond formation of compounds with structure G28 with the desired amine (HNR^NR⁷) give compounds with structure G29. Methods to form such amides will be apparent to those skilled in the art, but include for example the use of reagents such as HATU, HBTU, T3P and EDCI/HOBt, and the use of activated forms of the carboxylic acid such as the corresponding acyl halide, mixed anhydride or /V-hydroxysuccinimide ester. Reduction of the amide to the corresponding amine G30 will be apparent to those skilled in the art but such methods include but are not limited to the use of a reducing agent such as LiAIH._t. Suitable amine protecting groups and methods for the removal of said protecting groups will be known to those skilled in the art (for example Greene's Protective Groups in Organic Synthesis, 4th Edition). Upon deprotection, intermediates G31 can be converted to the desired compound, G32, by the procedure outlined in Scheme 1A.

Alternatively where X=H in G28, a reductive amination could be performed, for example using sodium triacetoxy borohydride and acetic acid to form G30 directly, rather than proceeding via intermediate G29.

G31 G32

Scheme 9A

General synthesis method 10

Scheme 10A illustrates the addition of an amine (HNR⁸R⁹) as a substituent which is a part of A. This can be achieved by coupling a relevant carboxyiic acid to a primary amine or a secondary amine, NHR⁸R⁹. Methods to form such amides will be apparent to those skilled in the art, but include for example the use of reagents such as HATU, HBTU, T3P and

EDCI/HOBt, and the use of activated forms of the carboxyiic acid such as the corresponding acyl halide, mixed anhydride or /V-hydroxysuccinimide ester. The group denoted by (X) may be but not limited to halogen, tosylate or other suitable group. Conversion of (X) in G33 into an ester in G34 will be apparent to those skilled in the art, but include for example a carbonylation reaction which can be achieve by the use of carbon monoxide in the presence of an transition metal catalyst such as but not limited to PdC dppf.DCM; and an alcoholic solvent such as but not limited to methanol, ethanol, isopropanol or iert-butyl alcohol.

Formation of the carboxyiic acid can be achieved by for example hydrolysis with a base such as an alkali metal hydroxide or an acid for example aqueous hydrochloric acid to form G35. The amide formation to form G36 can be achieved by the methods outline in Scheme 1A.

G35 G36

Scheme 10A

Alternatively, for the synthesis of ester G34 the order of steps can be reversed as described in Scheme 10B.

G34

Scheme 10B

Alternatively for the synthesis of amide G36 the steps may be reordered such that the formation of the R⁸R⁹N amide on the A substituent occurs after the coupling of A to the amine G26. This may be achieved by coupling a suitable amine with an intermediate where A bears a suitable functional group for coupling, for example but not limited to a carboxylic acid or alkali metal carboxylate salt, as shown in Scheme 10C

G36

Scheme 10C General synthesis method 1 1

Scheme 1 1 A illustrates the addition of an R^{1 1} group, as a substituent which is part of A. This can be achieved using any suitable coupling reaction known to the person skilled in the art, for example by Suzuki coupling, Negishi coupling, SwAr reaction, Ullman coupling, Chan Lam coupling, an alkylation reaction e.g. SN2 or SN1 , a Mitsonobu reaction, a C-H activation reaction, a Kumada coupling , a Buchwald reaction etc. The groups denoted by R X and B¹ are chosen to be suitable for the coupling reaction employed. For example, in the case of a Suzuki coupling reaction (X) may be a halogen, tosylate or other suitable group and B¹ represents a suitable boron compound including, but not limited to, a boronic acid or boronic ester.

G35

G36

Scheme 11A

Examples of B¹ that can be used in the Suzuki coupling include, but are not limited to, those shown below.

The types of R¹¹X compounds that can be used in the Suzuki coupling include, but are not limited to, those shown in Table 1 . Table 1 :

In addition to scheme 1 1 A, the position of the (X) and (B¹) can be reversed as shown below in scheme 1 1 B. to give the same final compound G36. Similarly to Scheme 1 1 A, the groups denoted by R^{1 1}B¹ and (X) are chosen to be suitable for the coupling reaction employed. For example, in the case of a Suzuki coupling reaction (X) may be a halogen, tosylate or other suitable group and R¹¹B¹ represents a suitable boron compound including, but not limited to, a boronic acid of boronic ester.

G37 G36

Scheme 11B

The types of R¹¹B¹ compounds that can be used in the Suzuki coupling include, but are not limited to, those shown in Table 2.

Table 2:

A variety of coupling reactions may be used to introduce the R^{1 1} group other than Suzuki coupling, such as for example transition metal catalysed coupling reactions of for example tin (Stille type reaction) and zinc (Negishi type reaction) compounds. Substitution of the halogen by suitable nucleophiles in the presence or absence of other reagents such as for example transition metal compounds is also suitable.

Coupling reactions can also be used to prepare the carboxylic acids used above for amide formations, scheme 1 1 C. In the starting material G39 and G41 , A as described herein, consists of -A³X and -A³B¹ respectively. In the product G42, A as described herein, consists of -A³R¹¹. The groups denoted by (X) and B¹ are chosen to be suitable for the coupling reaction employed. Potentially applicable reactions include, but are not limited to: Suzuki coupling, Negishi coupling, SwAr reaction, Ullman coupling, Chan Lam coupling, an alkylation reaction e.g. SN2 or SN1 , a Mitsonobu reaction, a C-H activation reaction, a Kumada coupling, a Buchwald reaction etc. For example, in the case of a Suzuki coupling reaction (X) may be a halogen, tosylate or other suitable group and B¹ represents a suitable boron compound including, but not limited to, a boronic acid of boronic ester.

Scheme 11 C

In G39 and G41 R¹² can be a H or a carbon group for example but not limited to Me, Et, Pr, iPr, Bu, t-Bu. It may be necessary to form the carboxylic acid before use in the amide coupling (Scheme 1 A), generally this can be achieved by for example hydrolysis with a base such as an alkali metal hydroxide or an acid for example aqueous hydrochloric acid to form G42. The same method for converting an ester to a carboxylic acid is used in other general schemes.

General synthesis method 12

Scheme 12A illustrates the addition of an R¹³ group, as a substituent which is part of A. This can be achieved using any suitable coupling reaction known to the person skilled in the art, for example, by a SwAr displacement or Buchwald coupling. The group denoted by (X) may be but not limited to halogen and is chosen to be suitable for the coupling reaction employed.

G43 G44 G45

Scheme 12A

In G43 and G44 R¹⁴ can be a H or a carbon group for example but not limited to Me, Et, Pr, iPr, Bu, t-Bu. In these instances in may be necessary to form the carboxylic acid before use in an amide coupling (Scheme 1A), generally this can be achieved by, for example, hydrolysis with a base such as an alkali metal hydroxide or an acid, for example, aqueous hydrochloric acid to form G45. The same method for converting an ester to a carboxylic acid is used in other general schemes. Alternatively, to synthesise ether linked compounds, a similar strategy can be employed as shown in Scheme 12B. This can be achieved using any suitable coupling reaction known to a person skilled in the art, for example, by a SNAr displacement, an Ullman-type coupling or a Chan Lam coupling to give compounds with structure G46. Upon hydrolysis using methods previously described, compounds with structure G47 may be obtained and used in an amide bond formation as shown in scheme 1A.

G43 G46 G47

Scheme 12B

Throughout the above Scheme suitable protecting groups may be required on R^4a or R^4b. In the case of either substituent being an alcohol, a silyl protecting group could be used, e.g. TBS. In the case of an amino substituent, suitable protecting groups could include Boc, phthalimide or dimethoxybenzyl,

If R^4a is OH in G36 it can be transformed through a range of functional group

interconversions that are known to those skilled in the art. For example R^4a = NH2 can be introduced using a reagents such as DPPA and DBU to give the azide, which can then be reduced using triphenylphosphine. R^4a = CN can be introduced via activation of the alcohol to a suitable leaving group, e.g. R^4a = mesylate, tosylate or halogen and then treatment with a suitable source of cyanide, e.g. sodium cyanide. The resulting cyano compound can be hydrolysed to the primary amide R^4a = CONH2 using acidic or basic conditions. Under more forcing acid or basic conditions, the carboxylic acid could be formed, i.e. R^4a = CO2H. The carboxylic acid could be reduced to the corresponding primary alcohol, R^4a = CH2OH using a suitable reducing agent, e.g. BH3, or alternatively through activation of the acid to a mixed anhydride, R^4a = CO2COR, where R = alkyl, and then treatment with a suitable reducing agent, e.g. sodium borohydride.

Further Embodiments

n

In some embodiments, n is 1 . In some embodiments, n is 2.

R^N

In some embodiment, R^N is H. In other embodiments, R^N is Me. R¹

In some embodiments, there may be no R¹ substituents. In some embodiments, R¹ represents one to four Me or halo groups, preferably one to three Me or halo groups and more preferably one or two Me or halo groups. In some of these embodiments, R¹ may represent F. In others of these embodiments, R¹ may represent Me groups.

R²

R^2a, R^2b, R^2c and R^2d (if present) are independently selected from H, F, CH₂OH and Me. In some of these embodiments, R^2a, R^2b, R^2c and R^2d (if present) are independently selected from H, Me and CH₂OH. In further of these embodiments, R^2a, R , R^2c and R^2d (if present) are independently selected from H and Me.

In some embodiments R^2a, R , R^2c and R^2d (if present) are all H.

In some of those embodiments where R^2a, R^2b, R^2c and R^2d are all present (i.e. formulae la and lb), they may be comprised of three H and one Me or CH2OH group. It may be preferred in these embodiments that R^2a is Me and R² , R^2c and R^2d are H. It may be preferred in these embodiments that R^2c is Me or CH2OH and R^2a, R^2b and R^2d are H.

In some of those embodiments where R^2a, R^2b, R^2c and R^2d are all present (i.e. formulae la and lb), they may be comprised of two H and two Me groups. It may be preferred in these embodiments that R^2a and R ^c are Me and R^2b and R^2d are H. It may be preferred in these embodiments that R^2a and R² are Me and R^2c and R^2d are H. It may also be preferred in these embodiments that R^2c and R^2d are Me and R^2a and R ^b are H.

In some of those embodiment where only R^2a and R^2b are present (i.e. formula Ic), they may be comprised of one H and one Me or CH2OH group. In some of those embodiment where only R^2a and R^2b are present (i.e. formula Ic), they may both be a Me or CH2OH group.

R³

R^3a and R^3b are independently selected from H and Me. In some embodiments R^3a is H and R^3b is Me. In some embodiments R^3a and R^3b are both H. In some embodiments R^3a and R^3b are both Me. _R4a

In some embodiments R^4a is OH. In other embodiments, R^4a is -Nh½. In other embodiments R^4a is -C(=0)N H2. In other embodiments R^4a is -CH₂OH.

It may be preferred that R^4a is OH. R*^b

In some embodiments R^4b is H. In some embodiments R^4b is Me.

R⁵

In some embodiments R⁵ is H. In some embodiments R⁵ is Me.

Enantiomers

The carbon to which R^4a and R^4b are attached is a chiral centre.

When the compound contains this chiral centre, in some embodiments, the compound is a racemate.

When the compound contains this chiral centre, in some embodiments, the compound is a single enantiomer. In some of these embodiments, the compound is the ff? -enantiomer. In others of these embodiments, the compound is the fS -enantiomer.

The compound may also include further chiral centres, for example, in compounds of formula la, in tetrahydronaphthalenyl group. In some embodiments, the compound is a racemate. In other embodiments, the compound is a single enantiomer. In some of these embodiments, the compound is the ff -enantiomer. In others of these embodiments, the compound is the (S -enantiomer.

R'-R⁵ & n

In some embodiments, R^N, R¹, R^2a, R^2b, R^2c, R^2d, R^3a, R^3b, R^4b and R⁵ are all H, R^4a is OH and n is 1 , and thus the compound of formula la is of formula la 1 , the compound of formula lb is of formula Ib1 , and the compound of formula lc is of formula Id :

In some embodiments, R\ R^2a, R^2b, R^2c, R^2d, R^3a, R³ , R^4b and R⁵ are all H, R^4a is OH and n is 2, and thus the compound of formula la is of formula Ia2, the compound of formula lb is of formula Ib2, and the com ound of formula Ic is of formula Ic2:

A

Optional substituents

When the optional substituent on A is Ci-₄ alkyl, it may be preferably selected from methyl, ethyl, APr, f-Bu.

When the optional substituent on A is Ci-₄ fluoroalkyl, it may preferably be selected from -CF₃ and -CF₂H. When the optional substituent on A is C5-6 heteroaryl, it may be substituted by one or more C1-4 a Iky I groups. These groups may preferably be on one or more of the nitrogen ring atoms (if present). These groups may also preferably be methyl.

When the optional substituent on A is C5-6 heteroaryl, it may preferably be selected from pyridizinyl, pyrimidinyl, pyridinyl, pyrazolyl, pyrazinyl, oxadiazolyl, isoxazolyl, triazolyl, imidazolyl, benzimidazolyl and thiadiazolyl. When the optional substituent on A is C5-6 heteroaryl methyl, it may preferably be selected from -CH₂-imidazolyl and -CH₂-triazolyl.

When the optional substituent on A is C5-6 heterocyclyl, it may preferably be morpholino. When the optional substituent on A is C5-6 heterocyclyl methyl, it may preferably be selected from -Ch -morpholino and -Ch -piperazinyl.

When the optional substituent on A is phenyl, it may be substituted by one or more C1-1 a Iky I groups. These groups may preferably be methyl.

When the optional substituent on A is phenyl, it may be substituted by one or more C1-4 fluoroalkyl groups. These groups may preferably be trifluoromethyl.

When the optional substituent on A is phenyl, it may be substituted by one or more C1-1 alkoxy groups. These groups may preferably be methoxy.

When the optional substituent on A is phenyl, it may be substituted by one or more halo substituents. These groups may preferably be fluoro or chloro, more preferably fluoro. When the optional substituent on A is phenyl, it may be substituted by one or more cyano groups. It may be preferred that there is a single cyano substituent.

When the optional substituent on A is halo, it may preferably be selected from F, CI and Br. When the optional substituent on A is amido, the amido substituent groups R and R' may preferably form a ring, which ring may also be bridged or substituted. If the amido group is not cyclic, it may preferably be selected from -C(=0)NH2, -C(=0)NMeH, -C(=0)NMe2 and - C(=0)N'PrH. If the amindo group is cyclic, it may preferably be selected from -C(=0)- piperidinyl, -C(=0)-hydroxypiperidinyl, -C(=0)-methoxypiperidinyl, -C(=0)-pyrrolidinyl,- C(=0)-morpholino, -C(=0)-methylmorpholino, -C(=0)-dimethylmorpholino and -C(=0)- azetidinyl. Further cyclic amido groups include:

When the optional substituent on A is amidomethyl, the amido substituent groups R and R' may preferably form a ring, which ring may also be bridged or substituted. If the amido group is not cyclic, the amidomethyl group may preferably be selected from

-CH₂C(=0)NMeH and -CH₂C(=0)N'PrH. If the amido group is cyclic, the amidomethyl group may preferably be selected from -CH2C(=0)-pyrrolidinyl -CH2C(=0)-morpholino, -C(=0)- hydroxypiperidinyl, -C(=0)-methoxypiperidinyl, -C(=0)-methylmorpholino and CH2C(=0)- azetidinyl . Further cyclic amidomethyl groups include:

When the optional substituent on A is acylamido, it may preferably be γ-lactam.

When the optional substituent on A is acylamidomethyl, it may preferably be selected from

-CH₂NHC(=0)Me, -CH₂NHC(=0)CF₃.

When the optional substituent on A is Ci-4 a Iky I ester, it may preferably be -C(=0)-OMe.

When the optional substituent on A is Ci-4 a Iky I ester methyl, it may preferably be

-CH₂-C(=0)-OMe.

When the optional substituent on A is Ci-4 alkyl carbamoyl methyl, it may preferably be -CH₂NHC(=0)OMe.

When the optional substituent on A is Ci-4 alkylacyl, it may preferably be selected from -C(=0)Me and -C(=0)Et. When the optional substituent on A is Ci-4 alkylacylmethyl, it may preferably be

-CH₂C(=0)Me.

When the optional substituent on A is phenylcarbonyl, it may preferably be -C(=0)-Ph. When the optional substituent on A is ether, it may preferably be selected from methoxy, ethoxy, -OBn, -OPh, -OCF₃,-OCF₂H, -0-(C₆H₄)-CN, -O-oxanyl, -OCH₂pyridinyl,

-OCH₂.oxadiazolyl, -OCH₂-isoxazole, When the optional substituent on A is amino, the amino substituent may be a C->-6 heteroaryl group, in which case the amino group may preferably be selected from -NH-pyrazinyl, -NH- pyrimidine. In other embodiments, the amino substituent may be a C₄-6 heterocyclyl group, such as optionally N-substituted azetidinyl, optionally N-substituted piperidinyl and oxetanyl. A further amino group may be:

When the optional substituent on A is aminomethyl, it may preferably be -CH2NH2.

Alternatively, the amino substituent may be as defined above

When the optional substituent on A is sulfonamido it may preferably be selected from -S0₂NMePh, -S0₂NMe₂, and -S0₂NHEt.

When the optional substituent on A is sulfonamino, it may preferably be selected from -NHS0₂Ph and -NHS0₂Me.

When the optional substituent on A is sulfone, it may preferably be -S0₂CF₃.

Optionally substituted phenyl

In some embodiments A may be an optionally substituted phenyl.

In some of these embodiments, A is unsubstituted phenyl.

In some of these embodiments, the phenyl of A has 1 , 2. 3. 4 or 5 substituents. In some of these embodiments, the phenyl of A has 1 or 2 substituents. It may be preferred that in some of these embodiments R¹, R^2a, R^2b, R^2c, R^2d, R^3a, R^3b, R^4b and R⁵ are all hydrogen and that R^4a is OH and n is 1.

It may be preferred in some of these embodiments that at least one of R¹, R^2a, R^2b, R^2c, R^2d, R^3a, R^3b, R^4b and R⁵ is not hydrogen.

It may be preferred in some of these some of these embodiments that R^1"5 and n is such that the compound is of formula la , Ia2, Ib1 , Ib2, Id or Ic2. It may be preferred in these embodiments that the optional substituents are independently selected from the following: C alkyl; Ci-₄ fluoroalkyl; C3-6 cycloalkyl; C5-6 heteroaryl; C5-6 heteroaryl methyl; C₄-e heterocyclyl; C₄-6 heterocyclyl methyl; phenyl; benzyl; halo; amido; amidomethyl; acylamido; acylamidomethyl; C alkyl ester; C alkyl ester methyl; Ci_-4 alkyl carbamoyl; CM alkyl carbamoyl methyl; CM alkylacyl; CM alkyl acyl methyl; phneylcarbonyl; carboxy; carboxymethyl; ether; amino; aminomethyl; sulfonamido; sulfonamino; sulfone; nitrile; and nitrilemethyl.

It may be preferred in these embodiments that the optional substituents are selected from: CM alkyl, fluoro, chloro, bromo, acetyl, methoxy, ethoxy, -C(=0)Me, -C(=0)Et, - CH₂C(=0)Me, phenyl, -CF₃, -CF₂H. -CN, -CH₂CN, -OBn, -OPh, -OCF3.-OCF2H. -0-(C₆H₄)- CN, -COOH, -CH2COOH, -C(=0)OMe, -C(=0)NH₂, -C(=0)NMeH, -C(=0)NMe₂, - C(=0)N'PrH, -C(=0)-piperidinyl, -C(=0)-pyrrolidinyl,-C(=0)-morpholino (which may be bridged or substituted by one, two or three methyl groups), -C(=0)-azetidinyl, - CH₂C(=0)NH₂, -CH₂C(=0)-azetidinyl, -CH₂C(=0)NMeH, -CH₂C(=0)N'PrH, -CH₂C(=0)- pyrrolidinyl, -CH₂C(=0)-morpholino, -CH₂-morpholino, -CH₂-methylpiperazinyl, -

OCH₂pyridinyl, -OCH₂-methyloxadiazolyl, -CH₂-imidazolyl, -O-tetrahydropyranyl, -CH₂- tetraydropyanyl,-NH-methylpyrazinyl, -CH₂-triazolyl, -NHS0₂Ph, -NHS0₂Me, -S0 NMePh, -S0₂NMe₂, -S0₂NHEt, -S0₂CF₃,- γ-lactam. -CH₂NHC(=0)Me, -CH₂NHC(=0)OMe. - CH₂NHC(=0)CF₃, morpholino, -CH₂NH₂, -C(=0)Ph, -OCH₂-isoxazolyl, -NH-pyrimidinyl, pyridizinyl, pyrimidinyl, pyridinyl, pyrazolyl, methylpyrazolyl, dimethylpyrazolyl, pyrazinyl, pyridazinyl , methyloxadiazolyl, oxadiazolyl, dimethyloxadiazolyl, isoxazolyl, dimethyltriazolyl, imidazolyl, benzimidazolyl and thiadiazolyl.

It may be preferred in these embodiments that, when the optional substituent is a C5-6 heteroaryl group, the heteroaryl ring itself is substituted with one or more CM alkyl groups. It may be preferred in the above embodiments that 1 substituent is present. In may be preferred in the above embodiments that 2 substituents are present.

Halo and methoxy (including CF3O) substituents may be preferred in the ortho position of the phenyl group. Ethoxy and a Iky I (e.g. methyl, CF2H and CF3) substituents may also be preferred in the ortho position of the phenyl group. Alkyl and C5-6 heteroaryl may be preferred in the meta position of the phenyl group. Amido and amidomethyl substituents may be preferred in the para position of the phenyl group. Particular favoured groups in the ortho position are ethoxy, methoxy, CI, F and CF₂H.

A combination of subsitutents that may be preferred is an ethoxy group in the ortho position of the phenyl group and an amido or amidomethyl group in the para position of the phenyl group. In some embodiments, the phenyl group bears a halo or methoxy substituent in the ortho position, and an amido or amidomethyl substituent in the para position of the phenyl group.

In some embodiments, the phenyl group bears an amino substituent in the meta position. Where the substituent on phenyl is a fused C5-6 Ni-containing heterocyclic ring, A may have a core structure selected from:

Particular A groups of interest include:

Optionally substituted naohthyl

When A is naphthyl, it may be in any orientation, e.g. naphth-1 -yl, naphth-2-yl.

In some embodiments A may be optionally substituted naphthyl.

In some of these embodiments, A is unsubstituted naphthyl. In some of these embodiments, the naphthyl ring of A has 1 , 2, 3. 4 or 5 substituents.

In some of these embodiments, the naphthyl ring of A has 1 or 2 substituents.

It may be preferred that in some of these embodiments R\ R^2A, R² , R^2C, R^2D, R^3A, R³ , R^4B and R⁵ are all hydrogen and that R^4A is OH and n is 1.

It may be preferred in some of these embodiments that at least one of R¹, R^2A, R^2B, R^2c, R^2D, R^3a, R³ , R^4B and R⁵ is not hydrogen. It may be preferred in some of these some of these embodiments that R^{1 "5} and n is such that the compound is of formula la , Ia2, Ia3, Ia2, Ib2 or Ic2.

It may be preferred in these embodiments that the optional substituents refers to 0-2 substituents independently selected from the following: CM alkyl; C fluoroalkyl; C3-6 cycloaikyl; C5-6 heteroaryl; C5-6 heteroaryl methyl; C4-6 heterocyclyl; C4-6 heterocyclyl methyl; phenyl; benzyl; halo; amido; amidomethyl; acylamido; acylamidomethyl; CM alkyl ester; CM a Iky I ester methyl; C1-4 alkyl carbamoyl; C1-4 alkyl carbamoyl methyl; C1-4 alkylacyl; C1-4 a Iky I acyl methyl; phneylcarbonyl; carboxy; carboxymethyl; ether; amino; aminomethyl;

sulfonamido; sulfonamino; sulfone; nitrile; and nitrilemethyl.

It may be preferred in these embodiments that 1 substituent is present. In may be preferred in these embodiments that 2 substituents are present.

Optionally substituted &>. ,? heteroarvl

In some embodiments A may be an optionally substituted C5-12 heteroaryl group.

In some of these embodiments, A is unsubstituted C5-12 heteroaryl group.

In some of these embodiments, the C5-12 heteroaryl of A has 1 , 2, 3, 4 or 5 substituents.

In some of these embodiments, the C5-12 heteroaryl of A has 1 or 2 substituents.

It may be preferred in these embodiments that the C5-12 heteroaryl ring is selected from one of the following: pyridinyl, pyrimidinyl, pyrazinyl, isoxazolyl, oxazolyl, thiophenyl, thiazolyl, thiadiazolyl, pyrazolyl, pyridonyl, imidazolyl, benzimidazolyl, imidazopyridinyl and quinolinyl. The heteroatoms may be in any location in the ring, which may be joined to the remainder of the molecule via a ring carbon atom. It may be further preferred that the C5-12 heteroaryl ring is either pyridinyl or pyrimidinyl. It may also be further preferred that the C5-12 heteroaryl is selected from pyridyl, pyrimidinyl, oxazolyl, oxadiazolyl, pyrazolyl and thiazolyl and in particular:

oxazol-4-yl

4- ridyl 2-pyridyl pyrimidin-4-yl

1 ,3,4-oxadiazol-2-yl 1 H-pyrazol-4-yl thiazol-5-y!

Further preferred groups may include benzothiazolyl and benzimidazolyol and in particular:

1 H-benzimidazol-4-yl

1 ,3-benzoihiazol-4-yl

It may be preferred that in some of these embodiments R¹, R^2a, R^2b, R^2c, R^2d, R^3a, R^3b, R^4b and R⁵ are all hydrogen and that R^4a is OH and n is 1.

It may be preferred in some of these embodiments that at least one of R¹, R^2a, R^2b, R^2c, R^2d, R^3a, R^3b, R⁴ and R⁵ is not hydrogen.

It may be preferred in some of these some of these embodiments that R^1"5 and n is such that the compound is of formula la , Ia2, Ib1 , Ib2, Id or Ic2.

It may be preferred in these embodiments that the optional substituents are independently selected from the following: CM alkyl; C1-4 fluoroalkyl; C3-6 cycloalkyl; C5-6 heteroaryl; C5-6 heteroaryl methyl; C4-6 heterocyclyl; C4-6 heterocyclyl methyl; phenyl; benzyl; halo; amido; amidomethyl; acylamido; acylamidomethyl; Cu alkyl ester; C alkyl ester methyl; CM alkyl carbamoyl; C1-4 alkyl carbamoyl methyl; C1-4 alkylacyl; C alkyl acyl methyl; phneylcarbonyl; carboxy; carboxymethyl; ether; amino; aminomethyl; sulfonamido; sulfonamino; sulfone; nitrile; and nitrilemethyl. It may be preferred in these embodiments that the optional substituents are selected from: CM alkyl; CM fluoroalkyl; C5-6 heteroaryl, C4-6 heterocyclyl; phenyl; halo; and ether.

It may be preferred in these embodiments that the optional substituent are selected from; methyl, ethyl, butyl, chloro, bromo, fluoro, morpholino, pyrrolidinyl, -OBn, -OPh, phenyl, para- bromophenyl, pyrazolyl, pyrimidinyl, imidazolyl and -CF3.

In may be preferred in these embodiments that 1 substituent is present. In may be preferred in these embodiments that 2 substituents are present. Halo and methoxy substituents may be preferred in the ortho position of a Ce heteroayl group, or a-position of C5 and C7-12 heteroaryl group. Amido and amidomethyl substituents may be preferred in the para position of a Ce heteroayl group, or γ-position of C5 and C7-12 hetereoaryl group.

In some embodiments, a Ce heteroayl group bears a halo or methoxy substituent in the ortho position, and an amido or amidomethyl substituent in the para position.

In some embodiments, a Ce heteroayl group bears an amino substituent in the meta position. In some embodiment, a C5 or C7-12 heteroaryl group bears an amino substituent in the β-position.

Where the Ce heteroayl group is 4-pyridyl, it may bear an ether substituent, for example in the 3-position. In some of these embodiments, the ether substituent may be -O-C4-6 heterocyclyl, wherein the Ct-e heterocyclyl may itself bear an ester group (e.g. methoxy ester).

Where the Ce heteroayl group is 2-pyridyl, it may bear an amido substituent, for example in the 4-position.

Where the Ce heteroayl group is 4-pyrimidinyl, it may bear an amino substituent, for example in the 3-position. In some of these embodiments, the amino substituent may be -NH- C₄-e heterocyclyl, wherein the Ct-e heterocyclyl may itself bear an acyl group (e.g. -C(=0)Me). Where A is a C5 heteoraryl group (e.g. oxazolyl, oxadiazolyl, pyrazolyl and thiazolyl), it may bear an amino, phenyl or Ce heteroaryl substituent in the β-position.

In some embodiments A is selected from one of the following groups:

63

n some embodiments A may be selected from one of the following groups:

In some embodiments A may be selected from one of the following groups:

In some embodiments, A may be selected from: phenyl, with a para-amido substituent and an optional ortho-ethoxy substituent; phenyl, with a para-ether substituent; 6-amino-3- pyridyl; 5-amino-3-pyridyl; and 2-amino-4-pyridyl.

It may be preferred that in some of these embodiments R^N, R¹, R^2a, R^2b, R^2c, R^2d, R^3a, R³ , R^4b and R⁵ are all hydrogen and that R^4a is OH and n is 1.

EXAMPLES

The following examples are provided solely to illustrate the present invention and are not intended to limit the scope of the invention, as described herein.

Acronyms

For convenience, many chemical moieties are represented using well known abbreviations, including but not limited to, methyl (Me), ethyl (Et), n-propyl (nPr), isopropyl (iPr), n-butyl (nBu), ferf-butyl (tBu), phenyl (Ph), benzyl (Bn), methoxy (MeO), ethoxy (EtO), trimethylsilyl

(TMS), and acetyl (Ac).

For convenience, many chemical compounds are represented using well known

abbreviations, including but not limited to, methanol (MeOH), deuterated methanol (£¾- MeOD) ethanol (EtOH), isopropanol (/-PrOH), ether or diethyl ether (Et₂0), ethyl acetate (EtOAc), acetic acid (AcOH), acetonitrile (MeCN), dichloromethane (methylene chloride, DCM), trifluoroacetic acid (TFA), dimethylformamide (DMF), tetrahydrofuran (THF), dimethylsulfoxide (DMSO), deuterated chloroform (CDC ), diethylamine (DEA), deuterated dimethylsulfoxide (cfe-DMSO), A/-ethyl-/V'-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI.HCI, EDCI), meia-chloroperoxybenzoic acid (mCPBA), 1 ,1 '- bis(diphenylphosphino)ferrocene (dppf), iert-butyloxycarbonyl (Boc, BOC), 2- (trimethylsilyl)ethoxymethyl (SEM), triethylamine (E iN or TEA), 2-(1 H-7-azabenzotriazol-1- yl)-1 , 1 ,3,3-tetramethyluronium hexafluorophosphate (HATU), 4-dimethylaminopyridine (DMAP), A/,A/-diisopropylethylamine (DIPEA or DIEA), 1 , 1 '-bis(diphenylphosphino)ferrocene dichloropalladium (II) (PdC (dppf)), irans-dichlorobis(triphenylphosphine)palladium(ll) (PdCI₂(PPh₃)2), tris(dibenzylideneacetone) dipalladium(O) (Pd₂(dba)₃),

tetrakis(triphenylphosphine)palladium(0) (Pd(PPh3)₄), propylphosphonic anhydride (T3P), hexamethylphosphoramide (HMPA), 1 ,2-dichloroethane (DCE), benzyl (Bn) and 1 - hydroxybenzotriazole (HOBt).

In addition, TLC refers to thin layer chromatography.

General Experimental Details

Unless otherwise stated the following generalisations apply. ¹H NMR spectra were recorded on a Bruker Ultrashield Plus (400 MHz), a Bruker AVANCE (400 MHz) or a Bruker Avance DRX300 (300 MHz). The multiplicity of a signal is designated by the following abbreviations: s, singlet; d, doublet; t, triplet; q, quartet; dd, doublet of doublets; dt, doublet of triplets; tt, triplet of triplets; br, broad; m, multiplet. All observed coupling constants, J, are reported in Hertz.

LCMS data was generated using either an Agilent 6100 Series Single Quad LCMS (LCMS- A). an Agilent 1260 Infinity Series UPLC/MS (LCMS-B), a Agilent 1200 Series G61 10A Quadrupole LCMS (LCMS-C), a Waters 2695 alliance (LCMS-D) or a Finnigan instrument (LCMS-E). Chlorine isotopes are reported as ³⁵CI, Bromine isotopes are reported as either ⁷⁹Br or ⁸¹ Br or both ⁷⁹Br/⁸¹ Br. LCMS Method A (LCMS-A):

Instrument: Agilent 6100 Series Single Quad LC/MS

Agilent 1200 Series HPLC

Pump: 1200 Series G131 1 A Quaternary pump

Autosampler: 1200 Series G1329A Thermostatted Autosampler Detector: 1200 Series G1314B Variable Wavelength Detector

LC conditions:

Reverse Phase HPLC analysis

Column: Luna C8 (2) 5 μιτι 50 * 4.6 mm 100 A

Column temperature: 30 C

Injection Volume: 5 L

Solvent A: Water 0.1 % Formic Acid

Solvent B: MeCN 0.1 % Formic Acid

Gradient: 5-100 % solvent B over 10 min

Detection: 254 nm or 214 nm

MS conditions:

Ion Source: Quadrupole

Ion Mode: Multimode-ES

Drying gas temp: 300 °C

Vaporizer temperature: 200 C

Capillary voltage (V): 2000 (positive)

Capillary voltage (V): 4000 (negative)

Scan Range: 100-1000

Step size: 0.1 sec

Acquisition time: 10 min

LCMS Method B (LCMS-B):

Instrument: Agilent 1260 Infinity Series UPLC/MS

Pump: 1260 Infinity G1312B Binary pump

Autosampler: 1260 Infinity G1367E 1260 HiP ALS

Detector: 1290 Infinity G4212A 1290 DAD

LC conditions:

Reverse Phase HPLC analysis Column: Poroshell 120 EC-C18 2.7 μπι 50 ^χ 3.0 mm

Column temperature: 35 °C

Injection Volume: 1 μΙ_

Solvent A: Water 0.1 % Formic Acid

Solvent B: MeCN 0.1 % Formic Acid

Gradient: 5-100 % solvent B over 3.8 min

Detection: monitored at 254 nm and 214 nm

MS conditions:

Ion Source: Quadrupole

Ion Mode: API-ES

Drying gas temp: 350 C

Capillary voltage (V): 3000 (positive)

Capillary voltage (V): 3000 (negative)

Scan Range: 100-1000

Step size: 0.1 sec

Acquisition time: 5 min

LCMS Method C (LCMS-C):

Instrument: Agilent 1200 Series G61 10A Quadrupole

Pump: Binary pump

Detector: DAD

LC conditions:

Reverse Phase HPLC analysis

Column: Xbridge-C18, 2.5 μπι, 2.1 *30 mm

Column temperature: 30 °C

Injection Volume: 1-10 μΙ_

Solvent A: Water 0.07 % Formic acid

Solvent B: Methanol

Gradient: 30-95 % solvent B over 3.5 min (for medium polarity samples) or 10-95% solvent B over 3.7 min (for large polarity samples)

Detection: monitored at 254 nm and 214 nm MS conditions:

Ion Source: Quadrupole Ion Mode: ES+

Drying gas temp: 350 C

Drying gas flow: 10 L/min

Nebulizer pressure: 35 psi

Capillary voltage (V): 3500 (positive) Scan Range: 50-900

LCMS Method D (LCMS-D)

Instrument: Waters ZQ 3100 Mass Detector Waters 2545-Pump

Waters SFO System Fluidics Organizer Waters 2996 Diode Array Detector

Waters 2767 Sample Manager LC conditions:

Reverse Phase HPLC analysis

Column: XBridge TM C18 5 pm 4.6 * 100 mm Injection Volume: 10 pi- Solvent A: Water 0.1 % Formic Acid

Solvent B: MeCN 0.1 % Formic Acid

Gradient: 10-100 % B over 10 min

Flow rate: 1.5 m L/min

Detection: 100-600 nm

MS conditions

Ion Source: Single-quadrupol

Ion Mode: ES positive

Source Temp: 150 °C

Desolvation Temp: 350 °C

Detection: Ion counting

Capillary (KV): 3.00

Cone (V): 30

Extractor (V): 3

RF Lens (V): 0.1

Scan Range: 100-1000 Amu

Scan Time: 0.5 sec Acquisition time: 10 min

Gas Flow: 100 L/h

Desolvation: 650 L/h LCMS Method E (LCMS-E)

Instrument: Finnigan LCG Advantage Max Finnigan Surveyor LC Pump

Finnigan Surveyor Autosampler

Finnigan Surveyor PDA Detector

LC conditions:

Reverse Phase HPLC analysis

Column: Gemini 3 pm C18 20 * 4.0 mm 1 10 A Injection Volume: 10 [it

Solvent A: Water 0.1 % Formic Acid

Solvent B: MeCN 0.1 % Formic Acid

Gradient: 10-100 % B over 10 min

Detection: 100-600 nm MS conditions

Ion Source: Ion trap

Ion Mode: ES positive

Temp: 300 °C

Detection: Ion counting

Scan Range: 80-1000 Amu

Scan Time: 0.2 sec

Acquisition time: 10 min

Preparative mass-directed LC

Instrument:

Waters ZQ 3100 -Mass Detector

Waters 2545-Pump

Waters SFO System Fluidics Organizer Waters 2996 Diode Array Detector

Waters 2767 Sample Manager LC conditions:

Reverse Phase HPLC analysis

Column: XBridge TM C18 5 μπι 19 x 50 mm

Injection Volume 500 μΙ_

Solvent A: Water 0.1 % Formic Acid

Solvent B:MeCN 0.1 % Formic Acid

Gradient: 25-100 % B over 10 min

Flow rate: 19 mL/min

Detection: 100-600 nm

MS conditions:

Ion Source: Single-quadrupole

Ion Mode: ES positive

Source Temp: 150 °C

Desolvation Temp: 350 C

Detection: Ion counting

Capillary (KV)-3.00

Cone (V): 30

Extractor (V): 3

RF Lens (V): 0.1

Scan Range: 100-1000 Amu

Scan Time: 0.5 sec

Acquisition time: 10 min

Gas Flow

Desolvation L hour-650

Cone L/hour-100

Preparative RP-HPLC:

Agilent 1260 Infinity HPLC system

UV detection at 210 nm and 254 nm

Gradient or isocratic elution through a Phenomenex Luna C8 (2) column 100 A Axia (250 * 21 .2 mm; particle size 5 μ ι)

Flow rate: 10 mL/min

Gradients are as specified in the individual examples. Analytical thin-layer chromatography was performed on Merck silica gel 60 F254 aluminium- backed plates which were visualised using fluorescence quenching under UV light or a basic KMn0.i dip or Ninhydrin dip. Preparative thin-layer chromatography (prep TLC) was performed using Tklst (China), grand grade: (HPTLC): 8±2 pm>80 %; (TLC): 10-40 μπι. Type: GF254. Compounds were visualised by UV (254 nm).

Flash chromatography was performed using a Biotage I solera purification system using either Grace or RediSep® silica cartridges.

Column chromatography was performed using Tklst (China), grand grade, 100-200 meshes silica gel.

Microwave irradiation was achieved using a CEM Explorer SP Microwave Reactor.

Where necessary, anhydrous solvents were purchased from Sigma-Aldrich or dried using conventional methods.

Additional Cartridges used are as follows:

Phase Separator:

Manufacturer: Biotage

Product: ISOLUTE ® Phase Separator (3 mL unless otherwise stated)

SCX and SCX-2 cartridges:

Manufacturer: Biotage

Product: ISOLUTE ® SCX 1 g, (6 mL SPE Column unless otherwise stated)

Manufacturer: Biotage

Product: ISOLUTE ® SCX-2 1 g (6 mL Column)

Manufacturer: Silicycle

Product: SCX-2 500mg or 5g

Manufacturer: Agilent

Product: Bond Elut® SCX 10g Sample extraction cartridge:

Manufacturer: Waters

Product: Oasis ® HLB 35 cc (6 g) LP extraction cartridge Intermediate Preparations

(\) 1 -Amino-3-((2.3-dihvdro-1 H-inden-2-yl)amino)propan-2-ol (12)

(a) 2-(3-((2,3-Dihydro-1H-inden-2-yl)amino)-2-hydroxypropyl)isoindoline- 1,3-dione (11) A mixture of V-glycidylp thalimide (2.00 g, 9.84 mmol) and 2-aminoindane (1 .57 g, 1 1 .8 mmol) in ethanol (50 mL) was stirred at 80 °C for 24 hours. The volatiles were removed in vacuo and the residue was purified by column chromatography (2 ^χ 40 g S1O2 cartridges, 0- 20% MeOH (containing 1 % v/v 2 M NH₃ in MeOH) in DCM) then (40 g Si0₂ cartridge, 0-10% MeOH (containing 1 % v/v 2 M NH3 in MeOH) in DCM) to give the desired compound as a brown solid (1 .69 g, 51 %). Ή NMR (400 MHz, CDCI3) δ 7.87 - 7.81 (m, 2H), 7.74 - 7.69 (m, 2H), 7.20 - 7.15 (m, 2H), 7.15 - 7.1 1 (m, 2H), 3.99 - 3.92 (m, 1 H), 3.84 - 3.70 (m, 2H), 3.64 - 3.55 (m, 1 H), 3.18 - 3.10 (m, 2H), 2.82 (dd, J = 12.2, 3.8 Hz, 1 H), 2.75 (dd, J = 15.6, 6.1 Hz, 2H), 2.65 (dd. J = 12.2, 7.3 Hz, 1 H).

(b) 1 -Amino-3-((2, 3-dihydro- 1 H -inden-2-yl)amino)propan-2-ol (12)

A solution of 2-(3-((2, 3-dihydro- 1 /-/-inden-2-yl)amino)-2-hydroxypropyl)isoindoline-1 ,3-dione 11 (1 .69 g, 5.02 mmol) in EtOH (35 mL) and DCM (35 mL) was treated with hydrazine hydrate (50-60%, 0.94 mL, 15.0 mmol) and the mixture was stirred for 96 hours at room temperature under nitrogen. The suspension was filtered through Celite and the filtrate solvent was concentrated in vacuo. The residue was dissolved in MeOH and loaded onto an SCX cartridge (Agilent HF Bond Elut-SCX, 10 g cartridge). The cartridge was washed with methanol (5 column volumes) and the product was eluted with 2 M NH3 in MeOH (5 column volumes). The solvent was removed in vacuo to give the desired product as a brown oil (1 .00 g, 97%). ¹H NMR (400 MHz, CDCI₃) δ 7.23 - 7.17 (m, 2H), 7.16 - 7.1 1 (m, 2H), 3.69 - 3.57 (m, 2H), 3.21 - 3.1 1 (m, 2H), 2.85 - 2.72 (m, 4H), 2.68 - 2.54 (m, 2H). (ii) (S)-1 -Amino-3-((2.3-dihvdro-1 H-inden-2-yl)amino)propan-2-ol (14)

(a) (R)-2-(3-((2, 3-Dihydro- 1 H -inden-2-yl)amino)-2-hydroxypropyl)isoindoline- 1.3-dione (13) (S)- /V-giycidylphthalimide (2.20 g, 10.8 mmol) and 2-aminoindane (1 .70 g, 13.0 mmo!) were dissolved in ethanol (45 mL) and the reaction was heated under nitrogen at 80 C for 22 hours. The volatiles were removed under reduced pressure and the resulting dark residue was purified by column chromatography (40 g S1O2, 0-20% methanol/DCM) to give the desired compound (1 .80 g, 51 %); ¹ H MR (300 MHz, CDCI₃) 7.84 - 7.77 (m, 2H), 7.71 - 7.64 (m, 2H), 7.20 - 7.06 (m, 4H), 4.01 (m, 1 H), 3.84 - 3.68 (m, 2H), 3.64 - 3.55 (m, 1 H), 3.18 - 3.10 (m, 2H), 2.85 - 2.77 (m, 2H), 2.74 - 2.61 (m, 2H ).

(b) (S)- 1 -Amino-3-((2, 3-dihydro- 1 H-inden-2-yl)amino)propan-2-ol (14)

A mixture of (f?)-2-(3-((2,3-dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)isoindoline-1.3- dione 13(1 .84 g, 5.47 mmol) and hydrazine monohydrate (380 L, 7.67 mmol) in ethanol (16 mL) and DCM (16 mL) was stirred at room temperature for 20 hours. The resulting solid was filtered off through Celite, washed with DCM (20 mL) and concentrated under reduced pressure. The residue was dissolved in methanol (15 mL) and the solution applied evenly to SCX cartridges (3 x 5 g). The cartridges were washed with methanol (20 mL for each cartridge) and eluted with a 2 M solution of ammonia in methanol (20 mL for each cartridge). After concentration of the combined ammonia eluates, the resulting compound was subjected to another round of purification by SCX cartridge, as described, to give the desired compound (950 mg. 84%); ¹ H NMR (300 MHz, CDCI3) 7.18 - 7.09 (m, 4H ), 3.62 (m, 2H), 3.15 - 3.09 (m, 2H), 2.76 - 2.66 (m, 4H), 2.62 - 2.51 (m, 2H).

(iii) (ffl-1 -Amino-3-((2,3-dihvdro-1 H-inden-2-vl)amino)propan-2-ol (16)

(a) (S)-2-(3-((2,3-Dihydro-1H-inden-2-yl)amino)-2-hydroxypropyl)isoindoline- 1,3-dione (15) (f?)-N-Glycidylphihalimide (2.50 g, 12.30 mmol) and 2-aminoindane (1 .97 g, 14.7 mmol) were dissolved in ethanol (50 mL) and the reaction was heated under nitrogen at 80 C for 72 hours. The ethanol was removed under reduced pressure and the dark residue was purified by column chromatography (40 g S1O2, 0-10% methanol/DCM) to give the desired compound (1.22 g, 29%).

(b) (R)- 1 -Amino-3-((2,3-dihydro- 1 H-inden-2-yl)amino)propan-2-ol (16)

A mixture of (S)-2-(3-((2.3-dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)isoindoline-1 ,3- dione 15(1 .22 g, 3.63 mmol) and hydrazine monohydrate (252 pL, 5.09 mmol) in ethanol (13 mL) and DCM (13 mL) was stirred at room temperature for 20 hours. The resulting solid was filtered through Celite and washed with ethanol (20 mL). The volatiles were removed under reduced pressure. The residue was dissolved in methanol (15 mL) and the solution applied evenly to SCX cartridges (3 x 5 g). The cartridges were washed with methanol (20 mL) and eluted with a 2 M solution of ammonia in methanol (20 mL). The combined ammonia eluates were concentrated in vacuo to give the desired compound (740 mg, 99%).

(iv) 4-((1 -Methyl-1 H-pvrazol-4-vl)oxy)benzoic acid I7)

5-Hydroxy-1 -methylpyrazole (0.250 g, 2.55 mmol) was added portion-wise to a stirring suspension of methyl 4-fluorobenzoate (0.35 mL, 2.7 mmol) and potassium carbonate (0.70 g, 5.1 mmol) in DMSO (10 mL) at room temperature. The resulting suspension was heated at 150 °C overnight. Water (50 mL) was added and the resulting solution was stirred for 4 hours. The reaction was cooled to room temperature, water (50 mL) was added and extracted with EtOAc (3 * 30 mL). The organic layers were discarded. The aqueous layer was acidified with 6 M HCI (2 mL) and extracted with EtOAc (3 * 30 mL). The combined organic extracts were washed with brine (3 * 30 mL), dried (Na2S0₄), filtered and concentrated in vacuo to give a pale yellow oil. The oil was purified by column

chromatography (24 g Si0₂ cartridge. 50-100% EtOAc (with 1 % v/v AcOH) in petroleum benzine 40-60 °C) to give the desired compound as a colourless solid (327 mg, 59%). ¹ H MR (400 MHz, cfe-DMSO) δ 7.95 - 7.86 (m, 2H), 7.81 (s, 1 H), 7.42 (s, 1 H), 7.09 - 7.00 (m, 2H), 3.82 (s, 3H), LCMS-B RT 2.98 min; m/z 219.1 [M+H]⁺; 217.1 [M-H]\ (v) ffl)-1-Amino-3-((2,3-dihvdro-1 H-inden-2-yl)(2.4-dimethoxybenzyl)amino propan-2-ol (110)

n o

(a) N-(2.4-dimethoxybenzyl)-2.3-dihydro- 1H-inden-2-amine 19

To a solution of 2.3-dihydro-1 H-inden-2-amine (20.0 g, 150.2 mmol) In ethanol (300 mL) was added 2,4-dimethoxybenzaldehyde(25.0 g, 150.2 mmol). The reaction mixture was stirred at 70 °C for 2 h, then the reaction cooled to 0 °C and NaBH₄ (1 1 .4 g, 300.3 mmol) was added. The resulting mixture was stirred at 0 °C for 30 min and poured into water (150 mL), CH2CI2 (150 mL) was added and the resulting mixture filtered. The filtrate was concentrated under reduced pressure and the aqueous mixture extracted with EtOAc (50 mL x 3). The organic layer was washed with brine (80 mL x 2), dried (Na2S0₄) and concentrated to give the desired compound as a yellow oil (41 g, 96%): LCMS: RT 1 .45 min; m/z 284.2 [M+H]\

(b) (R)- 1 -Amino-3-((2,3-dihydro- 1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino)propan-2-ol 110 To a solution of A/-(2.4-dimethoxybenzyl)-2,3-dihydro-1 H-inden-2-amine I9 (30.0 g, 105.9 mmol) in ethanol (400 mL) was added (f?)-2-(oxiran-2-ylmethyl)isoindoline-1 ,3-dione (22.6 g, 1 1 1 .1 mmol). The resulting mixture was heated at reflux overnight then Ν2Η4Ή2Ο (15.9 g, 317.6 mmol) was added. The resulting mixture was heated at reflux a further 4 h, then the reaction cooled and the precipitate removed by filtration. The filter cake was rinsed with ethanol (100 mL x 2) and the filtrate concentrated under reduced pressure. The residue obtained was dissolved in CH2CI2 (200 mL), washed with saturated aqueous NaHCOa solution (80 mL x 3) and brine (80 mL x 3), dried (Na2SO.t) and concentrated to give the desired compound as a yellow oil (35.7 g, 95%): ¹ H NMR (400 MHz, Methanol-d₄) δ 7.21- 7.19 (d, 1 H), 7.15-7.14 (m, 2H), 7.09-7.07 (m, 2H). 6.53-6.47 (m, 2H), 3.82 (s, 3H), 3.79 (s, 3H), 3.78-3.75 (m. 1 H), 3.65-3.63 (m, 3H), 3.02-2.99 (m, 4H), 2.69-2.64 (m, 1 H),2.51 -2.42 (m, 3H); LCMS: RT 0.46 min; m/z 357.2 [M+H]\

(vi) (SV1 -Amino-3-((2.3-dihvdro-1 /+inden-2-vl)(2.4-dimethoxvbenzyl)amino)propan-2-ol (11 1 )

To a solution of /V-(2.4-dimethoxybenzyl)-2,3-dihydro-1 H-inden-2-amine I9 (10.0 g, 35.2 mmol) in ethanol (150 mL) was added (S)-2-(oxiran-2-ylmethyl)isoindoline-1.3-dione (7.5 g, 36.9 mmol). The resulting mixture was heated at reflux overnight then the reaction cooled and 2Η4Ή2Ο (4.9 g, 97.4 mmol) added. The resulting mixture was heated at reflux for a further 4 h, then filtered and the filter cake rinsed with ethanol (20 mL). The filtrate was concentrated and the residue obtained dissolved in CH2CI2 (200 mL), washed with saturated aqueous solution of NaHCOa (80 mL x 3) and brine (20 mL x 2), dried ( a2S0₄) and concentrated to give the desired compound as a yellow oil (1 1.4 g, 87%): ¹H NMR (400

MHz,Methanol-d₄) δ 7.21 -7.15 (m, 3H), 7.10-7.07 (m, 2H), 6.55-6.54 (m, 1 H), 6.50-6.48 (m, 1 H), 3.83 (s, 3H), 3.78 (s, 3 H), 3.80-3.70 (m, 2H), 3.63-3.61 (m, 2H), 3.03-2.95 (m, 4H), 2.83-2.60 (m, 1 H), 2.58-2.51 (m, 3H); LCMS: RT 0.52 min; m/z 357.2 [M+H]\ (vii) 1 -Amino-3-((2.3-dihvdro-1 H-inden-2-vl)(2.4-dimethoxvbenzvnamino propan-2-ol (112)

112 To a solution of A/-(2.4-dimethoxybenzyl)-2.3-dihydro-1 H-inden-2-amine I9 (10.0 g, 35.2 mmol) in ethanoi (150 mL) was added 2-(oxiran-2-ylmethyl)isoindoline-1 ,3-dione (7.5 g, 37.1 mmol). The resulting mixture was heated at reflux overnight, then cooled to room

temperature and Ν2Η4Ή2Ο (4.9 g, 97.4mmol) added. The resulting mixture was heated at reflux for a further 3h. The reaction was cooled to RT, filtered and the filter cake rinsed with ethanoi (50 mL). The filtrate was concentrated then dissolved in CH2CI2, washed with a saturated aqueous solution of NaHCO.3 (80 mL x 3) and brine (20 mL x 2), dried ( a2S0₄) and concentrated to give the desired compound as yellow oil (10.5 g, 83%): LCMS: RT 0.53 min; m/z 357.2 [M+H]\

(viii (ff)-4-((3-((2.3-Dihvdro-1 H-inden-2-vl)(2.4-dimethoxvbenzvl)amino)-2- hydroxypropyl)carton -benzoic acid (14)

(a) (R)-Methyl 4-((3-((2,3-dihydro- 1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino)-2- hydroxypropyl)carbamoyl)benzoate 113

To a solution of (f?)-1-amino-3-((2.3-dihydro-1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino) propan-2-ol 110 (25.0 g, 70.2 mmol) in CH2CI2 (500 mL) was added 4- (methoxycarbonyl)benzoic acid (13.9 g, 77.2 mmol), DIPEA (18.1 g, 140.4 mmol), HOBt (0.95 g, 7.0 mmol) and EDCI (16.2 g, 84.2 mmol). The resulting mixture was stirred at room temperature overnight. The reaction was partitioned with a saturated solution of NaHCOa and extracted with CH2CI2 (100 mL x 2). The combined organic layers were washed with brine (100 mL x 2), dried ( a2SO.i) and concentrated. The residue was purified by column chromatography (100% CH2CI2 to 1 % methanol in CH2CI2) to give the desired compound as a yellow oil (19.8 g, 55%): LCMS: RT 2.30 min; m/z 519.3 [M+H] \ (b) (R)-4-((3-((2,3-dihydro-1H-inden-2-yl)(2A-dimethoxybenzyl)amin

hydroxypropyl)carbamoyl)benzoic acid 114

To a solution of (ft)-methyl 4-((3-((2,3-dihydro-1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino) - 2-hydroxypropyi )-ca rba moyl )benzoate 113 (17.4 g, 33.7 mmoi) in a mixture of THF (250 mL) methanol (12 mL) and water (12 mL) was added LiOH H₂0 (7.0 g, 168.3 mmoi). The reaction was stirred at room temperature for 48 h. The solvent was removed and the residue diluted with water (200 mL). The pH of the mixture was adjusted to pH 6 by addition of 3 M aqueous HCI. The mixture was extracted with CH2CI2 (150 mL x 3). The combined organic layers were washed with brine (100 mL x 2), dried ( a2S0₄) and concentrated to give the desired compound as a white solid (9.4 g, 56%): LCMS: RT 2.34 min; m/z 505.3 [M+H]\ fix) 4-((3-((2.3-Dihvdro-1 H-inden-2-vn(2.4-dimethoxvbenzvl)amino)-2- hvdroxypropvDcarbamovDbenzoic acid (116)

byproduct

(a) Methyl 4-((3-((2, 3-dihydro- 1 H-inden-2-yl)(2, 4-dimethoxybenzyl)amino)-2- hydroxypropyl)carbamoyl)benzoate 115

To a solution of 4-(methoxycarbonyl)benzoic acid (1.6 g, 9.0 mmoi) in ChbC O OO mL) was added HATU (4.7 g, 12.3 mmoi) and DIPEA (2.2 g, 16.8 mmoi). The mixture was stirred at room temperature for 1 h then 1 -amino-3-((2,3-dihydro-1 H-inden-2-yl)(2,4- dimethoxybenzyl)amino)propan-2-ol 112 (4.0 g, 1 1 .2 mmol) was added. The resulting mixture was stirred at room temperature overnight. Water (100 mL) was added and the phases were separated. The organic layer was washed with a saturated aqueous solution of NaHCO (100 mL) and brine (100 mL), dried ( a2S0₄) and concentrated. The residue was purified by column chromatography (0.7% methanol in CH2CI2) to give the byproduct as a yellow oil (1 .4 g) further elution (4% methanol in CH2CI2) gave the desired compound as a yellow oil (2.8 g, 48%). LCMS: RT 2.33 min; m/z 519.3 [M+H]⁺. (b) 4-((3-((2,3-Dihydro- 1 H-inden-2-yl)(2, 4-dimethoxybenzyl)amino)-2-hydroxypropyl) carbamoylbenzoic acid 116

To a solution of (ft)-methyl 4-((3-((2,3-dihydro-1 /-/-inden-2-yl)(2.4-dimethoxybenzyl)amino) - 2-hydroxypropyl )-ca rba moyl )benzoate 115 (2.6 g, 5.0 mmol) in a mixture of THF (40 mL), methanol(2 mL) and water (2 mL) was added LiOH H₂0 (1 .1 g, 25.1 mmol). The resulting mixture was stirred at room temperature overnight. The solvent was removed and the residue obtained diluted with water (50 mL). The pH of the aqueous mixture was adjusted to 6 by addition of 4 M aqueous HCI solution, then extracted with CH2CI2 (100 mL x 3). The combined organic layers were washed with brine (50 mL), dried (Na2S0₄) and concentrated to give the desired compound as a white solid (1.6 g, 64%). LCMS: RT 2.19 min; m/z 505.3 [Μ+Η]·.

(x) 2-(4-((3-((2.3-Dihvdro-1 Wnden-2-vl)(2.4-dimethoxvbenzvnamino)-2- hvdroxvDropvDcarbamovl) phenvnacetic acid (118)

(a) Methyl 2-(4-((3-((2.3-dihydro- 1 H-inden-2-yl)(2, 4-dimethoxybenzyl)amino)-2- hydroxypropyl) carbamoyl)phenyl)acetate 117

To a solution of 4-(2-methoxy-2-oxoethyl)benzoic acid (1 .0 g, 5.1 mmol) in CH2C (50 mL) was added HATU (2.7 g, 7.1 mmol) and DIPEA (1.2 g, 9.7mmol). The mixture was stirred at room temperature for 1 h, then 1-amino-3-((2,3-dihydro-1 H-inden-2-yl)(2,4- dimethoxybenzyl)amino)propan-2-ol 112 (2.3 g, 6.4 mmol) was added. The resulting mixture was stirred at room temperature overnight, then diluted with water (80 mL). The organic layer was separated, washed with a saturated aqueous NaHCC solution (100ml_) , brine (100ml_), dried (Na2S04) and concentrated. The residue was purified by column

chromatography (0.5% methanol in CH2CI2) to give the desired compound as a yellow oil (0.92 g, 27%): LCMS: RT 2.24 min; m/z 533.4 [M+H]\

(b) 2-(4-((3-((2,3-Dihydro- 1H-inden-2-yl)(2,4-dimethoxybenzyl)amino)-2- hydroxypropyl)carbamoyl)phenyl)acetic acid 118

To a solution of methyl 2-(4-((3-((2,3-dihydro-1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino) -2- hydroxypropyl)carbamoyl)phenyl)acetate 117 (1.5 g, 2.8 mmol) in a mixture of THF (20 mL), methanol (2 mL) and water (2 mL) was added LiOH H2O (0.6 g, 1 .4 mmol). The resulting mixture was stirred at room temperature overnight. The solvent was removed, and the residue diluted with water (50 mL). The pH of the aqueous solution was adjusted to 6 by addition of 4 M aqueous HCI solution. The aqueous mixture was extracted with CH2CI2 (10 mL x 3) and the combined organic layers dried (Na2SO.i) and concentrated to give the desired compound as a yellow oil (0.63 g, 42%): LCMS: RT 2.16 min; m/z 519.3 [M+H]\

(xi) (ffl-4-((3-((2,3-Dihvdro-1 H-inden-2-yl (2.4-dimethoxybenzyl amino)-2- h droxyRroeYDcaitemovl)-3-ethoxvbe^^

(a) Ethyl-4-bromo-2-ethoxybenzoate 119

To a mixture of 4-bromo-2-hydroxybenzoic acid (20.0 g, 92.6 mmoi) and K2CO3 (38.4 g, 277.9 mmol) in dimethylsulfoxide (50 mL) at 40 C was added ethyl bromide (15.2 g, 138.9 mmol) dropwise over 30 min. The reaction was stirred for 2 h then further ethyl bromide (15.2 g, 138.9 mmol) was added over 30 min. The reaction was stirred a further 8 h then diluted with CH2CI2 (150 mL) and filtered. The filtrate was washed with water (200 mL x 10) and brine (200 mL x 3), dried over a2S0₄ and concentrated to give the desired compound as a brown solid (24.3 g, 96%): LCMS: RT 2.93 min; m/z 273.0 [M+H]\

(b) 4-Bromo-2-ethoxybenzoic acid 120

To a solution of ethyl 4-bromo-2-ethoxybenzoate 119 (24.1 g, 88.6 mmol) in a mixture of THF (150 mL), methanol (15 mL) and water (15 mL) was added LiOH H₂0 (18.6 g, 44.3 mmol). The resulting mixture was stirred at room temperature for 24 h. The solvent was removed, and the residue was diluted with water (200 mL). The pH of the aqueous mixture was adjusted to 6 by addition of 2 M aqueous HCI solution. The aqueous mixture was extracted with ΟΗ₂(¾ (150 mL x 3) and the combined organic layers washed with brine (100 mL x 3), dried (Na2S0₄) and concentrated to give the desired compound as a yellow solid (19.8 g, 92%): LCMS: RT 2.47 min; m/z 246.9 [M+H]⁺ for ⁸¹ Br.

(c) Benzyl- 4-bromo-2-ethoxybenzoate 121

To a solution of 4-bromo-2-ethoxybenzoic acid I20 (16.4 g, 67.2 mmol) in acetonitrile (75 mL) was added benzyl bromide (13.8 g, 80.7 mmol) and K2CO3 (18.6 g, 134.4 mmol). The resulting mixture was stirred at 40 °C overnight. The reaction mixture was filtered and concentrated. The residue obtained was diluted with CH2CI2 (50 mL) and washed with water (100mL x 2). The organic layer was washed with brine (70 mL x 3), dried (Na2S0₄) and concentrated. The residue was purified by column chromatography (1 % EtOAc in petroleum ether) to give the desired compound as a light yellow oil (20.9 g, 93%): LCMS: RT 3.31 min; m/z 334.9 [M+H]\

(d) 1 -Benzyl 4-methyl 2-ethoxyterephthalate 122

To a solution of benzyl 4-bromo-2-ethoxybenzoate 121 (20.0 g, 59.9 mmol) in methanol (100 mL) was added Pd(dppf)CI₂ (2.2 g, 3 mmol) and TEA (13.3 g, 131 .7 mmol). The resulting mixture was heated at reflux overnight under an atmosphere of carbon monoxide. The reaction mixture was concentrated and the residue obtained diluted with water and extracted with CH2CI2 (50 mL x 3). The combined organic layers were washed with brine (60 mL x 2), dried ( a2S0₄) and concentrated. The residue was purified by column chromatography (2% EtOAc in petroleum ether) to give the desired compound as a white solid (15.1 g, 81 %): LCMS: RT 3.03 min; m/z 315.1 [M+H]\

(e) 2-Ethoxy-4-(methoxycarbonyl)benzoic acid 123

To a solution of 1 -benzyl 4-methyl 2-ethoxyterephthalate I22 (14.9 g, 47.4 mmol) in THF (100 mL) was added 10% Pd/C (1 .5 g). The resulting mixture was stirred at room

temperature overnight under H2 atmosphere. The reaction mixture was filtered through celite, and the filter cake washed with THF (50mL). The filtrate was concentrated and the residue diluted with water. The pH of the aqueous solution was adjusted to 6 by addition of a 2 M aqueous HCI solution. The resultant mixture was extracted with CH2CI2 (50 mL x 3) and the combined organic layers washed with brine (50mL x 2), dried (Na₂SO.t) and concentrated to give the desired compound as a white solid (10.2 g, 96%): LCMS: RT 2.13 min; m/z 225.0 [M+H]\ (f) Methyl (R)-4-((3-((2.3-dihydro- 1 H-inden-2-yl)(2, 4-dimethoxybenzyl)amino)-2- hydroxypropyl) carbamoyl) -3-ethoxybenzoate 124

To a solution of 2-ethoxy-4-(methoxycarbonyl)benzoic acid 123 (3.8 g, 17.0 mmol) in CH2CI2 (40 mL) was added (fl)-1 -amino-3-((2, 3-dihydro- 1 H-inden-2-yl)(2,4- dimethoxybenzyl)amino)propan-2-ol 110 (5.5 g, 15.1 mmol), HOBt (208.6 mg.1.5mmol), DIPEA (4.0 g, 30.9 mmol) and EDCI (3.7 g, 18.5 mmol). The resulting mixture was stirred at room temperature overnight. The mixture was partitioned with saturated aqueous NaHCOa solution and extracted with ΟΗ₂(¾ (30 mL x 2). The combined organic layers were washed with brine (20 mL x 2), dried ( a2S0₄) and concentrated. The residue obtained was purified by column chromatography (1 % methanol in CH2CI2) to give the desired compound as a yellow oil (6.6 g, 76%): LCMS: T 2.41 min; m/z 563.2 [M+H]'.

(g) (R)-4-((3-((2,3-Dihydro-1H-inden-2-yl)(2 -dimethoxybenzyl)amino)-2- hydroxypropyl)carbamoyl) -3-ethoxybenzoic acid 125

To a solution of methyl (f?)-4-((3-((2,3-dihydro-1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino) - 2-hydroxypropyl)carbamoyl)-3-ethoxybenzoate I24 (6.5 g, 1 1.6 mmol) in a mixture of THF (40 mL), methanol (4 mL) and water (4 mL) was added LiOH H₂0 (2.4 g, 57.8 mmol). The resulting mixture was stirred at room temperature overnight. The mixture was concentrated, and the residue diluted with water (20 mL). The pH of the aqueous mixture was adjusted to 6 by addition of a 2 M aqueous HCI solution. The aqueous was then extracted with Ch C O O mL x 3) and the combined organic layers washed with brine (20 mL x 2), dried ( a2S0₄) and concentrated to give the desired compound as a white solid (6.3 g, 98%): LCMS: RT 2.26 min; m/z 549.3 [M+H]'.

(xii 4-(Morpholine-4-carbonyl)benzoic acid 027)

(a) Methyl 4-(morpholine-4-carbonyl)benzoate 126

To a solution of mono-methyl terephthalate (10.0 g, 55.9 mmol) in dichloromethane (500 mL) at 0 °C was added oxalyl chloride (8.5g, 66.6 mmol) and a catalytic amount of DMF (0.5 mL). The mixture was stirred at 0 °C for 3 h. Morpholine (9.7 g, 1 1 1.0 mmol) was added followed by triethylamine (7.9 g, 77.7 mmol), the reaction was allowed to warm to room temperature and stirred for 16 h. The reaction was diluted with a saturated aqueous solution of NaHCC (250 ml_). The aqueous layer was extracted with dichloromethane (3 * 250 mL) and the combined organic layers washed with 0.5 M aqueous HCI solution (100 mL), water (200 mL) and brine (100 mL). The organic layer was dried ( a2S04), filtered and

concentrated in vacuo to give a solid which was crystallized (petroleum ether: ethyl acetate=20:1 ) to give the desired compound (1 1 .3 g, 82%) as a yellow solid. LCMS: RT 1 .1 1 min; m/z 250.1 [M+H]⁺

(b) 4-(Morpholine-4-carbonyl)benzoic acid 127

To a solution of methyl 4-(morpholine-4-carbonyl)benzoate I26 (10.5 g, 42.1 mmol) in a mixture of THF (200 mL), MeOH (20 mL) and water (2 mL) was added LiOH.H₂0 (1 .9 g, 46.3 mmol) and the reaction stirred at room temperature for 24 h. The solvents were removed under reduced pressure and the resulting gum suspended in a 0.5 M aqueous citric acid solution (250 mL). The aqueous layer extracted with dichloromethane (3 * 250 mL) and the combined organic fractions dried (MgSC ) and concentrated in vacuo to yield the product as a tan solid (8.8 g, 88%). LCMS: RT 0.48 min; m/z 236.1 [M+H]\¹H NMR (400 MHz, Methanol^) δ = 8.23 - 8.04 (d, 2H), 7.67 - 7.48 (d, 2H), 3.79 (br s, 2H), 3.75 - 3.68 (m, 2H), 3.68 - 3.61 (m, 2H), 3.49 - 3.40 (m, 2H). (xiii) 3-(Morpholine-4-carbonyl)benzoic acid I29

I28 I29

(a) Methyl 3-(morpholine-4-carbonyl)benzoate 128

Morpholine (958 L, 1 1 .1 mmol, 1 equiv), mono-methyl isopthalate (2.00 g, 1 1 .1 mmol, 1 equiv), MeCN (50 mL), DIPEA (5.80 mL, 33.3 mmol, 3 equiv) and HATU (4.64 g, 12.2 mmol, 1.1 equiv) were stirred at room temperature. After two hours the mixture was quenched with 5% w/v aqueous sodium carbonate solution (50 mL) and the organic solvents removed in vacuo. The aqueous residue was extracted with ethyl acetate (3 * 50 mL), and the pooled organic extracts washed with water (2 x 50 mL), dried over sodium sulfate and concentrated. Chromatography (40 g silica cartridge, 0-50% ethyl acetate in petroleum benzine) and collection of the suspected product fractions gave the desired compound (2.72 g, 98% yield) as a pale brown oil. LCMS-B: RT= 3.32 min, m/z = 250 [M+H]\ (b) 3-(morpholine-4-carbonyl)benzoic acid 129

UOH.H2O (1.37 g, 32.7 mmol, 3 equiv) was added to a solution of I28 (2.72 g, 10.9 mmol) in MeOH (20 mL) and water (10 mL), the resulting suspension was stirred for 16 hours at room temperature. The volatiles were removed in vacuo to give a white solid. Water was added followed by a 0.5 M aqueous solution of citric acid until the solution was at pH 4. The mixture was stirred for 30 minutes before it was extracted with EtOAc (3 * 70 mL). The combined organic layers were washed with brine (100 mL), dried (MgSO.t) and concentrated in vacuo to give the desired compound (1.74 g, 68% yield) as a white solid. LCMS-B: RT= 3.19 min, m/z = 236 [M+H]⁺, 234 [M-H]\

(xiv) 2-(2-Cyclopropylacetyl)isoindoline-5-carboxylic acid 132

(a) 1-(5-Bromoisoindolin-2-yl)-2-cyclopropylethanone 130

To a solution of 2-cyclopropylacetic acid (96 mg, 0.96 mmol) in dichloromethane (10 mL) was added HOBt (212.8 mg, 1.39 mmol), DIPEA (421 .2 mg, 3.26 mmol) and EDCI (266.4 mg, 1.39 mmol). The mixture was stirred at 0 °C for 20 min then a solution of 5- bromoisoindoline (190 mg, 0.96 mmol) in dichloromethane (5 mL) was added. The resulting mixture was stirred at room temperature overnight. Water (20 mL) was added and the mixture extracted with dichloromethane (20 mL x 3). The combined organic layers were dried (Na2S0₄), filtered and concentrated in vacuo to give the crude product. The crude product was purified by column chromatography (Petroleum ether: ethyl acetate=10:1 ) to give the desired product (145 mg, 68%) as a yellow oil. LCMS: RT 2.69 min; m/z 280.2 [M+H]⁺. (b) Methyl 2-(2-cyclopropylacetyl)isoindoline-5-carboxylate 131

To a solution of 1-(5-bromoisoindolin-2-yl)-2-cyclopropylethanone I30 (140 mg, 0.5 mmol) in methanol (10 mL) was added triethylamine (1 1 1 .1 mg, 1.1 mmol) and Pd(dppf)C (18.3 mg, 0.025 mmol,). The mixture was heated at reflux under a carbon monoxide atmosphere overnight. The mixture was concentrated to give the crude product which was purified by column chromatography (Petroleum ether: ethyl acetate=10:1 ) to give the desired product (103 mg, 79%) as a yellow oil. LCMS: RT 2.38 min; m/z 260.3 [M+H]⁺

(c) 2-(2-Cyclopropylacetyl)isoindoline-5-carboxylic acid 132

To a solution of 2-(2-cyclopropylacetyl)isoindoline-5-carboxylic acid 131 (97 mg, 0.37 mmol) in mixture of THF (8 mL), methanol (0.8 mL), and water (0.08 mL) was added LiOH H₂0 (77.7 mg, 1.85 mmol). The resulting mixture was stirred at room temperature for 3 days. The mixture was concentrated to give the crude product which was dissolved in water and washed with dichloromethane (10 mL). The aqueous layer was neutralized by addition of a 0.5 M aqueous HCI solution (10 mL) and the solution extracted with dichloromethane (20 mL x 3). The combined organic layers were dried (NazSC^) and concentrated to give the desired product (50 mg, 55%) as a pale solid. LCMS: RT 2.1 1 min; m/z 246.3 [M+H] '

(xv) 2-(2-Cvclopropvlacetvn-1.2.3.4-tetrahvdroisoauinoline-7-carboxvlic acid I35

(a) 1 -(7-Bromo-3, 4-dihydroisoquinolin-2( 1 H)-yl)-2-cyclopropylethanone 133

To a solution of 2-cyclopropylacetic acid (201.2 mg, 2.01 mmol) in DCM (30 mL) was added DIPEA (1 .4 g, 8.8 mmol), HOBt (445.5 mg, 2.91 mmol) and EDCI (557.8 mg, 2.91 mmol). The mixture was stirred at 0 C for 20 min then 7-bromo-1.2,3,4-tetrahydroisoquinoline hydrochloride (500 mg, 2.01 mmol) was added. The resulting mixture was stirred at room temperature overnight. The reaction was diluted with water (20 mL) and the solution extracted with DCM (30 mL x 3). The combined organic layers were dried ( a2S0₄) and concentrated to give the crude product which was purified by column chromatography (petroleum ether: ethyl acetate =10:1 ) to give the desired product (420 mg, 71 %) as a yellow oil. LCMS: RT 2.59 min; m/z 294.0 [M+H]⁺

(b) Methyl 2-(2-cyclopropylacetyl)-1 ,2,3,4-tetrahydroisoquinoline-7-carboxylate 134

To a solution of 1-(7-bromo-3,4-dihydroisoquinolin-2(1 H)-yl)-2-cyclopropylethanone I33 (400 mg, 1.36 mmol) in MeOH (10 mL) was added triethylamine (303.0 mg, 3.0 mmol), and Pd(dppf)C (49.9 mg, 0.068 mmol). The mixture was heated at reflux under a carbon monoxide atmosphere overnight. The solvent was removed under reduced pressure to give the crude product which was purified by column chromatography (Petroleum ether: ethyl acetate=5:1 ) to give the desired product (160 mg, 79%) as a yellow oil. LCMS: RT 2.47 min; m/z 274.3 [M+H]⁺

(c) 2-(2-Cyclopropylacetyl)-1,2,3,4-tetrahydroisoquinoline-7-carboxylic acid 135

To a solution of methyl 2-(2-cyclopropylacetyl)-1 ,2,3.4-tetrahydroisoquinoline-7-carboxylate 134 (150 mg, 0.54 mmol) in a mixture of THF (8 mL), methanol (0.8 mL), and water (0.08 mL) was added LiOH H₂0 (69.3 mg, 1.65 mmol). The resulting mixture was stirred at room temperature for 2 days. The mixture was concentrated to give the crude product which was diluted with water and washed with dichloromethane (10 mL). The water layer was neutralized by addition of a 0.5 M aqueous HCI solution (10 mL) and extracted with dichloromethane (20 mL x 3). The combined organic layers were dried Na2S0₄) and concentrated to give the desired product (70 mg, 50%) as a pale solid. LCMS: RT 2.20 min; m/z 260.1 [M+H]⁺ fxvi 2-(2-Cvclopropvlacetvn-1.2.3.4-tetrahvdroisoauinoline-7-carboxvlic acid 138

I37 I38 (a) 1 -(6-Bromo-3, 4-dihydroisoquinolin-2( 1 H)-yl) -2-cyclopropylethanone 136

To a solution of 2-cyclopropylacetic acid (236.2 mg, 2.36 mmol) in DCM (30 mL) was added DIPEA (1 .03 g, 8.0 mmol), HOBt (523.6 mg, 3.42 mmol) and EDCI (655.6 mg, 3.42 mmol). The mixture was stirred at 0 °C for 20 min then 6-bromo-1.2,3,4-tetrahydroisoquinoline (500 mg, 2.01 mmol) was added. The resulting mixture was stirred at room temperature overnight. The reaction was diluted with water (20 mL) and the aqueous layer extracted with DCM (30 mL x 3). The combined organic layers were dried (Na^SC^) and concentrated to give the crude product which was purified by column chromatography (petroleum ether: ethyl acetate =10:1 ) to give the desired product (580 mg, 83%) as a yellow oil. LCMS: RT 2.69 min; m/z 294.0 [M+H]⁺

(b) Methyl 2-(2-cyclopropylacetyl)- 1,2.3, 4-tetrahydroisoquinoline-6-carboxylate 137

To a solution of 1-(6-bromo-3.4-dihydroisoquinolin-2(1 H)-yl)-2-cyclopropylethanone I36 (560 mg, 1.90 mmol) in MeOH (30 mL) was added triethylamine (422.2 mg, 4.18 mmol), and Pd(dppf)C (69.7 mg, 0.095 mmol). The mixture was heated at reflux under a carbon monoxide atmosphere overnight. The mixture was concentrated to give the crude product which was purified by column chromatography (Petroleum ether: ethyl acetate=5:1 ) to give the desired product (280 mg, 54%) as a pale oil. LCMS: RT 2.45 min; m/z 274.1 [M+H] ' (c) 2-(2-Cyclopropylacetyl)- 1,2,3,4-tetrahydroisoquinoline-7-carboxylic acid 138

To a solution of methyl 2-(2-cyclopropylacetyl)-1.2,3.4-tetrahydroisoquinoline-6-carboxylate I37 (200 mg, 0.73 mmol) in mixture of THF (10 mL), methanol (1 mL), and water (0.1 mL) was added LiOH H2O (94.0 mg, 2.26 mmol). The resulting mixture was stirred at room temperature for 2 days. The mixture was concentrated to give the crude product. Water was added and the aqueous layer washed with dichloromethane (10 mL). The aqueous layer was neutralized by addition of a 0.5 M aqueous HCI solution (10 mL) and the solution was extracted with dichloromethane (20 mL x 3). The organic layer was dried (NazSC ) and concentrated to give the desired product (170 mg, 89%) as a yellow solid. LCMS: RT 2.15 min; m/z 260.3 [M+H]⁺ (xyii 2-Acetvl-1.2,3.4-tetrahvdroisoquinoline-6-carboxylic acid 141

(a) 1 -(6-Bromo-3,4-dihydroisoquinolin-2(1 H)-yl)ethanone 139

To a solution of 6-bromo-1 ,2,3,4-tetrahydroisoquinoline (456.0 mg, 2.15 mmol) in DCM (20 mL) was added triethylamine (326.0 mg, 3.22 mmol) and acetic anhydride (219.5 mg, 2.15 mmol) at 0 °C. The resulting mixture was stirred at 0 °C for 3 h, then allowed to warm to room temperature and stirred overnight. The reaction mixture was washed with a saturated aqueous NaHC03 solution (20 mL x 2), the organic layer separated, dried (Na2S0₄) and concentrated to give the desired product (500 mg, 91 %) as an orange oil. LCMS: RT 2.41 min; m/z 254.0 [M+H]⁺

(b) Methyl 2-acetyl-1 ,2,3,4-tetrahydroisoquinoline-6-carboxylate 140

To a solution of 1-(6-bromo-3.4-dihydroisoquinolin-2(1 H)-yl)ethanone I39 (500 mg, 1.97 mmol) in MeOH (30 mL) was added triethylamine (437.0 mg, 4.33 mmol), and Pd(dppf)C (71.9 mg, 0.098 mmol). The mixture was heated at reflux under a carbon monoxide atmosphere overnight. The mixture was concentrated to give the crude product which was purified by column chromatography (Petroleum ether: ethyl acetate=2:1 ) to give the desired product (360 mg, 78%) as an orange oil. LCMS: RT 2.07 min; m/z 234.1 [M+H] ' (c) 2-Acetyl- 1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid 141

To a solution of methyl 2-acetyl-1 ,2,3,4-tetrahydroisoquinoline-6-carboxylate I40 (200 mg, 0.86 mmol) in a mixture of THF (10 mL), methanol (1 mL), and water (0.1 mL) was added LiOH H₂0 (108.3mg, 2.58 mmol). The resulting mixture was stirred at room temperature for 2 days. The mixture was concentrated to give the crude product which was diluted with water and washed with dichloromethane (10 mL). The aqueous layer was neutralized by addition of a 0.5 M aqueous HCI solution (10 mL) and the solution extracted with dichloromethane (20 mL x 3). The organic layer was dried (Na2SO.i) and concentrated to give the desired product (100 mg, 53%) as a white solid. LCMS: RT 3.01 min; m/z 220.1 [M+H] ' (xviii) 2-Acetvl-1.2.3.4-tetrahvdroisoguinoline-7-carboxvlic acid 144

(a) 1 -(7-Bromo-3, 4-dihydroisoquinolin-2( 1 H)-yl)ethanone 142

To a solution of 7-bromo-1 ,2,3.4-tetrahydroisoquinoline (500 mg, 2.0 mmol), triethylamine (650 mg, 6.4 mmol) and DMAP (25 mg, 0.2 mmol) in DCM (20 mL) at 0 °C was added acetic anhydride (210 mg, 2.1 mmol). The resulting mixture was stirred at room temperature overnight. The reaction was washed with NaHCC>3 solution (20 mL x 2) and the organic layer dried a2S0₄) and concentrated to give the desired product (350 mg, 70%) as a clear oil. LCMS: RT 3.03 min; m/z 254.1 [M+H]⁺

(b) Methyl 2-acetyl-1 ,2.3,4-tetrahydroisoquinoline-7-carboxylate 143

To a solution of 1-(7-bromo-3.4-dihydroisoquinolin-2(1 H)-yl)ethanone I42 (350 mg, 1.385 mmol) in MeOH (30 mL) was added triethylamine (307.1 mg, 2.25 mmol), and Pd(dppf)CI₂ (50.6 mg, 0.069 mmol). The mixture was heated at reflux under a carbon monoxide atmosphere overnight. The mixture was concentrated to give the crude product which was purified by column chromatography (Petroleum ether: ethyl acetate=3:1 ) to give the desired product (150 mg, 46%) as an orange oil. LCMS: RT 2.35 min; m/z 234.3 [M+H]' (c) 2-Acetyl- 1 , 2, 3,4-tetrahydroisoquinoline- 7-carboxylic acid 144

To a solution of methyl 2-acetyl-1 ,2,3,4-tetrahydroisoquinoline-7-carboxylate I43 (150 mg, 0.64 mmol) in a mixture of THF (10 mL), methanol (1 mL), and water (0.1 mL) was added LiOH H2O (80.6 mg, 1 .92 mmol). The resulting mixture was stirred at room temperature for 1 day. The mixture was concentrated to give the crude product which was diluted with water and washed with dichloromethane (10 mL). The water layer was neutralized by addition of a 0.5 M aqueous HCI solution (10 mL) and the solution was extracted with dichloromethane (20 mL x 3). The organic layer was dried Na2S0₄ and concentrated to give the desired product (80 mg, 57%) as a yellow oil. LCMS: RT 1 .57 min; m/z 220.1 [M+H]⁺

(xix) 6-(Piperidine-1 -carbonyl)nicotinic acid I47

I46 147

(a) (5-Bromopyridin-2-yl)(piperidin- 1 -yl)methanone 145

To a solution of 5-bromopicolinic acid (5.0 g, 25.0 mmol) in DCM (250 mL) was added HATU (1 1.4 g, 30.0 mmol) and Ε¾Ν (8.8 g, 88.0 mmol). The mixture was stirred at room

temperature for 30 min then piperidine (2.6 g, 30.0 mmol) was added and the resulting mixture stirred at room temperature overnight. Water (200 mL) was added and the organic layer separated. The aqueous layer was extracted with DCM (2 x 200 mL) and the combined organic fractions washed with brine (200 mL), dried ( a2S0₄) and concentrated. The residue was purified by column chromatography (0-30% EtOAc/petroleum ether) to give the desired compound as yellow oil (6.0 g, 90%): LCMS: RT 2.21 min, m/z 269.1 ; 271.1 [M+Hf for ^/9Br and ⁸¹ Br respectively

(b) Methyl 6-(piperidine- 1 -carbonyl)nicotinate 146

To a solution of (5-bromopyridin-2-yl)(piperidin-1 -yl)methanone I45 (4.0 g, 14.9 mmol) in MeOH (50 mL) was added Et₃N (3.3 g, 32.8 mmol) and PdC (dppf) (410 mg, 0.75 mmol). The resulting mixture was heated at reflux under a CO balloon overnight. The solvent was removed and the residue diluted with water (50 mL). The aqueous layer was extracted with DCM (3 x 50 mL) and the combined organic layers washed with brine (100 mL), dried ( a2S0₄) and concentrated. The residue was purified by column chromatography (0-30% EtOAc/petroleum ether) to give the desired compound as a yellow solid (650 mg, 18%): LCMS: RT 1 .86 min, m/z 249.1 [M+H]⁺ (c) 6-(Piperidine- 1 -carbonyl)nicotinic acid 147

To a solution of methyl 6-(piperidine-1 -carbonyl)nicotinate I46 (600 mg, 2.4 mmol) in MeOH (5 mL) was added a solution of NaOH (193 mg, 4.8 mmol) in water (0.5 mL). The resulting mixture was stirred at room temperature overnight. The solvent was removed and the residue resuspended in water. The pH of the aqueous solution was adjusted to pH 4 by addition of a 4 M aqueous HCI solution. The aqueous layer was extracted with DCM (5 x 50 mL) and the combined organic layers dried ( a2S0₄) and concentrated to give the desired compound as a yellow solid (300 mg, 53%): LCMS: RT 0.96 min, m/z 235.1 [M+H]' (xx) 2-Propoxybenzoic acid I49

(a) Propyl 2-propoxybenzoate 148

To a mixture of 2-hydroxybenzoic acid (5.0 g, 36.2 mmol), Nal (0.5 g, 3.6 mmol) and K2CO3 (13.4 g, 0.1 1 mol) in DMSO (10 mL) at 36 °C was added 1 -bromopropane (6.7 g, 54.3 mmol) dropwise over a period of 30 min. The reaction was stirred for 2 h, then further 1- bromopropane (6.7 g, 54.3 mmol) was added over a period of 30 min. The resulting mixture was stirred at 36 °C overnight then diluted with DCM (50 mL) and filtered. The solid was washed with DCM (2 x 50 mL) and the filtrate washed with water (5 x 100 mL) and brine (100 mL), dried (Na2S0₄) and concentrated to give the desired compound as colorless oil (7.4 g, 93%): ¹H NMR (400 MHz, CDCI₃) δ 7.78-7.76 (m, 1 H), 7.43-7.39 (m, 1 H), 6.96-6.92 (m, 2H), 4.27-4.24 (m, 2H), 4.00-3.96 (m, 2H), 1.89-1 .72 (m, 4H), 1.07-1.00 (m, 6H): LCMS: RT 2.26 min, m/z 223.1 [M+H]'

(b) 2-Propoxybenzoic acid 149

To a solution of propyl 2-propoxybenzoate 148 (1 .0 g, 4.5 mmol) in a mixture of THF (10 mL), MeOH (2 mL) and water (2 mL) was added LiOH.H₂0 (0.95 g, 22.5 mmol). The resulting mixture was stirred at room temperature overnight. The solvent was removed and the residue diluted with water (10 mL). The aqueous layer was washed with DCM (2 x 10 mL) and the pH adjusted to 2 by addition of a solution of concentrated HCI. The aqueous layer was extracted with DCM (3 x 10 mL) and the combined organic layers dried ( a2S0₄) and concentrated to give the desired compound as yellow oil (300 mg, 53%): ¹H NMR (400 MHz, CDCh) δ 10.98 (brs, 1 H), 8.19-8.17 (m, 1 H), 7.56-7.52 (m, 1 H), 7.14-7.10 (m, 1 H), 7.05-7.03 (m, 1 H), 4.23-4.20 (m, 2H), 1.99-1.90 (m, 2H), 1.12-1.08 (m, 3H): LCMS: RT 2.15 min, m/z 181.1 [M+H]^*

(xxjj2;lsopropoxybenzoic acid 151

150 151

(a) Isopropyl 2-isopropoxybenzoate 150

To a mixture of 2-hydroxybenzoic acid (5.0 g, 36.2 mmol), K2CO3 (13.4 g, 0.1 1 mol) and Nal (0.5 g, 3.6 mmol) in DMSO (10 mL) at 36 °C was added 2-bromopropane (6.7 g, 54.3 mmol) dropwise over a period of 30 min. The reaction was stirred for 2 h, then further 2- bromopropane (6.7 g, 54.3 mmol) was added dropwise over 30 min. The resulting mixture was stirred at 36 °C overnight. The mixture was diluted with DCM (50 mL) and the solids removed by filtration. The filter cake was washed with DCM (2 x 50 mL) and the combined filtrates washed with water (5 x 100 mL) and brine (100 mL), dried (Na2SO.t) and

concentrated to give the desired compound as a colorless oil (2.3 g, 29%): ¹H NMR (400 MHz, CDCh) δ 7.70-7.68 (m, 1 H), 7.40-7.36 (m, 1 H), 6.97-6.91 (m, 2H), 5.28-5.21 (m, 1 H), 4.61-4.55 (m, H), 1.36-1.34 (m, 12H): LCMS: RT 2.19 min, m/z 223.1 [M+H]^*

(b) 2-lsopropoxybenzoic acid 151

To a solution of propyl 2-isopropoxybenzoate I50 (1 .0 g, 4.5 mmol) in a mixture of THF (10 mL), /-PrOH (1 mL) and water (0.1 mL) was added LiOH.H₂0 (0.95 g, 22.5 mmol). The resulting mixture was stirred at room temperature for 3 days. The solvent was removed and the residue diluted with water (20 mL). The aqueous layer was washed with DCM (20 mL) and the pH adjusted to 2 by addition of concentrated HCI. The aqueous layer was extracted with DCM (3 x 20 mL) and the combined organic layers dried ( a2S0₄) and concentrated to give the desired compound as yellow oil: (600 mg, 75%): ¹H NMR (400 MHz, d₄-MeOD) δ 7.85-7.82 (m, 1 H), 7.47-7.43 (m, 1 H), 7.07-7.05 (m, 1 H), 6.99-6.96 (m, 1 H), 4.72-4.65 (t, 1 H), 1 .33-1 .31 (m, 6H): LCMS: RT 2.15 min, m/z 181.1 [M+H]^* (xxii) 2-(2.2.2-Trifluoroethoxy)benzoic acid 152

152

To a solution of 2,2,2-trifluoroethanol (4.8 g, 47.9 mmol) in DMF (30 mL) was added sodium ierf-butoxide (2.8 g, 28.7 mmol) at 0 °C. The solution was stirred at room temperature for 1 h then 2-chlorobenzoic acid (3.0 g, 19.2 mmol) and CuBr (247 mg, 1.7 mmol) were added. The reaction was heated at 120 °C for 5 hours under nitrogen, then 20% v/v aqueous HCI solution was added. The resulting mixture was extracted with CH2CI2 (60 mL) and the combined organic layers dried (Na2SO.i) and concentrated. The residue was purified by column chromatography (2% MeOH/ in CH2CI2) to give the crude product (1 .8 g). 100 mg of this material was further purified by prep TLC (EtOAc:petroleum ether = 1 :1 ) to give the desired compound (67 mg, 29%) as a white solid: Ή NMR (400 MHz, CD₃OD) δ 7.65-7.63 (m, 1 H), 7.51-7.47 (m, 1 H), 7.18-7.16 (m, 1 H), 7.10-7.06 (m, 1 H), 4.77-4.70 (m, 2 H); LCMS: RT 3.60 min; m/z 221.1 [M+H]⁺.

(xxiii) 4-((3 5ffl-3.5-Dimethvlmorpholine-4-carbonyl)benzoic acid I54

I53 |₅₄

(a) Methyl 4-((3R,5R)-3,5-dimethylmorpholine-4-carbonyl)benzoate 153

To a solution of 4-(methoxycarbonyl)benzoic acid (285 mg, 1.6 mmol) in CH2CI2 (10 mL) was added DIPEA (594 mg, 4.6 mmol) and HATU (608 mg, 1 .6 mmol) and the mixture stirred for 1 h. (3 ?,5f?)-3,5-dimethylmorpholine (200 mg, 1.3 mmol) was added and the reaction stirred at room temperature for 2 days. The mixture was diluted with water (50 mL) and extracted with CH2CI2 (3 x 30 mL). The combined organic layers were washed with brine (50 mL), dried ( a2S0₄) and concentrated. The residue was purified by column chromatography (20% EtOAc in petroleum ether) to give the desired compound as a white solid (200 mg, 55%): ¹H NMR (400 MHz, CD₃OD) δ 8.1 1-8.10 (m, 2 H), 7.61 -7.59 (m, 2 H), 3.93 (s, 3 H), 3.92-3.89 (m, 4 H), 3.53-3.49 (m, 2 H), 1.29- 1 .27(s, 6 H); LCMS : RT 2.22 min; m/z 278.1 [M+H]⁺.

(b) 4-((3R,5R)-3,5-Dimethylmorpholine-4-carbonyl)benzoic acid 154

To a solution of methyl 4-((3 5 ?)-3,5-dimethylmorpholine-4-carbonyl)benzoate 153 (180 mg, 0.65 mmol) in a mixture of THF (5 mL), MeOH (0.5 mL) and water (0.05 mL) was added UOH.H2O (100 mg, 2.34 mmol) and the resulting mixture stirred at room temperature overnight. The solvent was removed and the residue diluted with water (5 mL). The aqueous solution was acidified to pH 3 by addition of a 1 M aqueous HCI solution and extracted with CH2CI2 (3 x 50 mL). The combined organic extracts were dried (Na2SO.i) and concentrated to give the desired compound as a white solid (150 mg, 88%): ¹H NMR (400 MHz,CD₃OD) δ 8.12-8.10 (m, 2H), 7.60-7.58 (m, 2H), 3.93-3.89 (m, 4 H), 3.53-3.49 (m, 2 H), 1.29-1 .28 (m, 6 H); LCMS RT 1 .67 min; m/z 264.1 [M+H] \ (xxiv 2-Methoxv-4-(2-morpholino-2-oxoethyl)benzoic acid I59

(a) Methyl 2-methoxy-4-methylbenzoate 155

To a solution of 2-hydroxy-4-methylbenzoic acid (10.0 g, 66 mmol) in acetone (500 mL) was added K2CO3 (18.0 g, 131 mmol). Me2S0₄ (22 g, 178 mmol) was then added dropwise and the resulting mixture heated at reflux for 14 hours. The mixture was poured into cold water and extracted with EtOAc (70 mL x 3). The combined organic layers were dried (Na2SO.i), filtered and concentrated and the residue was purified by column chromatography (1 % EtOAc in petroleum ether) to give the desired compound as a yellow liquid (7.6 g, 64%). LCMS: RT 2.31 min; m/z 181.1 [M+1]\

(b) 2-(3-Methoxy-4-(methoxycarbonyl)phenyl)acetic acid 156

To a solution of diisopropylamine (1 .3 g, 13.3 mmol) in dry THF (50 mL) at -50 °C was added n-BuLi (2.5M in hexane, 5.4 mL, 13.3 mmol) dropwise under nitrogen atmosphere. The mixture was stirred at -50 °C for 30 min and then HMPA (2.4 g, 13.3 mmol) was added, followed by methyl 2-methoxy-4-methylbenzoate I55 (2.0 g, 1 1.1 mmol). The resultant mixture was stirred at -78 °C for 2 h then CO? gas was bubbled through the mixture. During the course of the addition, the reaction mixture became pale, the mixture was stirred for another 30 min then poured into a solution of 1 M aqueous HCI (100 mL). The aqueous layer was extracted with dichloromethane and the combined organic layers washed with water and brine, dried (Na2SO.t) and concentrated. The residue was purified by column

chromatography (0-1 % methanol in dichloromethane) to give the desired compound (270 mg, 1 1 %) as colorless oil. LCMS T 1 .50 min; m/z 225.1 [M+H]\

(c) Methyl 4-(2-chloro-2-oxoethyl)-2-methoxybenzoate 157

To a solution of 2-(3-methoxy-4-(methoxycarbonyl)phenyl)acetic acid 156 (135 mg, 0.6 mmol) in dichloromethane (5 mL) with a catalytic amount of DMF (1 mL) was added oxalyl chloride (99 mg, 0.8 mmol). The mixture was stirred at room temperature overnight then the reaction mixture was concentrated to give the desired product which was used for the next step without further purification.

(d) Methyl 2-methoxy-4-(2-morpholino-2-oxoethyl)benzoate 158

To a solution of methyl 4-(2-chloro-2-oxoethyl)-2-methoxybenzoate 157 (146 mg, 0.6 mmol) in dichloromethane (5 mL) was added DIPEA (155 mg, 1.2 mmol) and morpholine (63 mg, 0.7 mmol). The reaction was stirred at room temperature overnight then the reaction mixture was partitioned with water, washed with water (2x20 mL) and brine and concentrated to afford the crude product. The residue was purified by column chromatography (0-10% EtOAc in petroleum ether) to give the desired compound (67 mg, 38%) as a colorless oil. LCMS: RT 1 .16 min; m/z 294.1 [M+H]\

(e) 2-Methoxy-4-(2-morpholino-2-oxoethyl)benzoic acid 159

To a solution of Methyl 2-methoxy-4-(2-morpholino-2-oxoethyl)benzoate 158 (67 mg, 0.2 mmol) in a mixture of THF (3 mL), MeOH (0.3 mL) and water (0.3 mL) was added LiOH.H₂0 (19 mg, 0.5 mmol). The mixture was stirred at room temperature overnight then

concentrated under reduced pressure. The residue was dissolved in water and the pH adjusted to 2 by addition of a 1 M aqueous HCI solution. The aqueous mixture was extracted with dichloromethane (3 x 5 mL) and the combined organic layers dried (Na2S0₄) and concentrated to give the desired compound which was used for the next step without further purification. LCMS: RT 0.41 min; m/z 280.1 [M+H]\ (xxv) 1-Oxa-7-azaspiro[3.5lnonane I60

I60

Benzyl 1 -oxa-7-azaspiro[3.5]nonane-7-carboxylate(1 g, 3.8 mmol) and 10% Pd/C (100 mg) in methanol (10 mL) was stirred at room temperature under a hydrogen atmosphere overnight. The reaction mixture was filtered through celite and the filtrate concentrated. The residue was used for the next step without further purification. LCMS: RT 0.30 min; m/z 128.1 [M+H]\

(xxy 4-(2-Methoxv-2-oxoethyl)benzoic acid 161

161

To a solution of 4-(carboxymethyl)benzoic acid (5.0 g, 27.7 mmol) in methanol (60 mL) was added thionyl chloride (0.1 mL, 1.3 mmol). The reaction was stirred at room temperature overnight. The reaction mixture was concentrated and the residue was purified by chromatography (dichloromethane) to give the desired compound (3.0 g, 57%) as a white solid. LCMS: RT 2.00 min; m/z 195.1 [M+H]\

(xxvii 4-(2-Oxo-2-(pyrrolidin-1-vl)ethvl)benzoic acid I64

(a) 2-(4-Bromophenyl)- 1 -(pyrrolidin- 1 -yl)ethanone 162

To a solution of 2-(4-bromophenyl)acetic acid (2.0 g, 4.3 mmol) in THF (10 mL) was added HATU (3.6 g, 4.7 mmol), DIPEA (2.2 g, 8.6 mmol) and pyrrolidine (0.64 g, 4.5 mmol). The solution was stirred at room temperature overnight then poured into a 1 M aqueous solution of HCI (20 mL). The aqueous layer was extracted with dichloromethane, washed with water and brine, dried (Na2S0₄), concentrated and the residue obtained purified by column chromatography (0-2% methanol in dichloromethane) to give the desired compound (2.0 g, 84%) as a white solid.

(b) Methyl 4-(2-oxo-2-(pyrrolidin- 1 -yl)ethyl)benzoate 163

A solution of 2-(4-bromophenyl)-1-(pyrrolidin-1-yl)ethanone I62 (1.15g, 4.3 mmol),

Pd(dppf)C (157 mg, 0.2 mmol) and triethylamine (953 mg, 9.4 mmol) in methanol (60 mL) under a CO (g) atmosphere was heated at reflux overnight. The reaction mixture was concentrated and the residue was purified by column chromatography (dichloromethane) to give the desired compound (130 mg, 13%) as an orange oil. LCMS: RT 2.1 1 min; m/z 248.1 [M+H] \

(c) 4-(2-Oxo-2-(pyrrolidin- 1 -yl)ethyl)benzoic acid 164

To a solution of methyl 4-(2-oxo-2-(pyrrolidin-1 -yl)ethyl)benzoate I63 (130 mg, 0.53 mmol) in a mixture of THF (2 mL), methanol (0.2 mL) and water (0.2 mL) was added LiOH.H₂0 (27 mg, 0.63 mmol) and the mixture stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure, the residue was dissolved in water and then acidified to pH 4 by the addition of a 1 M aqueous HCI solution. The aqueous layer was extracted with dichloromethane (2 x 5 mL) and the combined organic fractions dried

(NazSC ) and concentrated to give the desired compound (67 mg, 54%) which was used for the next step without further purification. LCMS: RT 3.31 min; m/z 234.1 [M+H]\

(xxviii) 4-(2-Morpholino-2-oxoethyl)benzoic acid I67

(a) 2-(4-Bromophenyl)-1 -morpholinoethanone 165

To a solution of 2-(4-bromophenyl)acetic acid (500 mg, 2.3 mmol) in dichloromethane (50 mL) was added morpholine (243 mg, 2.8 mmol), HATU (1 .06 g, 2.8 mmol) and Et₃N (810 mg, 8.0 mmol). The reaction was stirred at room temperature overnight. Water was added and the organic phase separated, washed with water (100 mL) and brine, dried (NazSCi), concentrated and the residue purified by chromatography (0-50% EtOAc in petroleum ether) to give the desired compound (500 mg, 76%) as a yellow solid. LCMS: RT 2.21 min; m/z 284.0 [M+H]⁺. (b) Methyl 4-(2-morpholino-2-oxoethyl)benzoate 166

A solution of 2-(4-bromophenyl)-1-morpholinoethanone I65 (446 mg, 1 .6 mmol), Pd(dppf)Cb (62 mg, 0.08 mmol) and triethylamine (377 mg, 3.7 mmol) in methanol (60 mL) under a CO (g) atmosphere was heated at reflux overnight. The reaction mixture was concentrated and the residue was purified by column chromatography (0-10% EtOAc in petroleum ether) to give the desired compound (329 mg, 80%) as an orange oil. LCMS: RT 1.43 min; m/z 264.1 [M+H]⁺. (c) 4-(2-Morpholino-2-oxoethyl)benzoic acid I67

To a solution of methyl 4-(2-morpholino-2-oxoethyl)benzoate I66 (329 mg, 3.8 mmol) in a mixture of THF (10 mL), methanol (1 mL) and water (1 mL) was added LiOH.h O (319 mg, 7.6 mmol) and the mixture stirred at room temperature overnight. The reaction mixture was concentrated and the residue dissolved in water. The pH of the solution was adjusted to 4 by addition of a 1 M aqueous HCI solution and the aqueous mixture was extracted with dichloromethane (2 x 25 mL). The combined organic layers were dried (NazSO-t) and concentrated to give the desired compound (1 18 mg, 38%) as a yellow solid. LCMS: RT 2.67 min; m/z 250.1 [M+H]\ (xxix) 1 -Benzvl-2-oxo-1.2-dihvdropyridine-4-carboxvlic acid I68

To a suspension of NaH (60% dispersion in mineral oil, 1.1 g, 25.3 mmol) in THF (16 mL) at 0 °C was added 2-oxo-1 ,2-dihydropyridine-4-carboxylic acid (0.78 g, 5.6mmol). The reaction was stirred at 0°C for 30 min then benzyl bromide (1 .9 g, 1 1.2 mmol) and DMF (2 mL) were added. The resulting mixture was stirred at room temperature overnight, water (2 mL) was added and the pH of the aqueous layer adjusted to 4 by addition of a 3 M aqueous HCI solution. The aqueous layer was extracted with CH2CI2 CI O mL x 2) and the combined organic layers washed with brine (15 mL x 2), dried ( a2S0₄) and concentrated to give the desired compound as a yellow liquid (219.1 mg, 17%): LCMS: RT 1 .81 min; m/z 230.1

[M+H] '.

(a) Ethyl-2-ethoxy-4-methylbenzoate 169

To a mixture of 2-hydroxy-4-methylbenzoic acid (8.2 g, 53.9 mmoi) and K2CO3 (22.4 g, 161.7 mmol) in dimethylsulfoxide (70 mL) at 40 °C was added and ethyl iodide (12.6 g, 80.8 mmol) dropwise over a period of 30 min. The reaction was stirred for 2 h then further ethyl iodide (12.6 g, 80.8 mmol) was added over 30 min. The resulting mixture was stirred a further 8 h at 40 °C, then diluted with Ch C O SO mL) and filtered. The filtrate was washed with water (200 mL x 10) and brine (200 mL x 2), dried ( a2S0₄) and concentrated to give the desired compound as a yellow liquid (10.1 g, 90%): LCMS: RT 2.70min; m/z 209.1 [M+H]\

(b) 3-Ethoxy-4-(ethoxycarbonyl)-benzoic acid 170

To a solution of ethyl 2-ethoxy-4-methylbenzoate 169 (10.0 g, 48.1 mmol) in a mixture of pyridine (25 mL) and water (75 mL) was added KMn0.i (22.8g, 144.2 mmol). The resulting mixture was heated at 50 °C for 48 h, then cooled and allowed to stir at room temperature for 24 h. The mixture was filtered and the filter cake washed with hot water. The combined aqueous filtrates were washed with EtOAc (75 mL x 3) and acidified with 2M aqueous HCI solution. The mixture was extracted with CH2CI2 (150 mL x 3). The combined organic layers were washed with brine, dried (Na2S0₄) and concentrated to give the desired compound as a white solid (5.0 g, 44%): LCMS: RT 0.25 min; m/z 239.0 [M+H]\ (c) Ethyl 4-(3-oxa-8-azabicyclo[3.2.1]octane-8-carbonyl)-2-ethoxybenzoate 171

To a solution of 3-ethoxy-4-(ethoxycarbonyl)benzoic acid I70 (2.5 g, 10.4 mmol) in CH2CI2 (20 mL) was added 3-oxa-8-azabicyclo[3.2.1]octane hydrochloride (1 .42 g, 9.5 mmol), HOBt (135.1 mg, 1.0 mmol), DIPEA (2.5 g, 19.0 mmol) and EDCI (2.2 g, 1 1 .4 mmol). The resulting mixture was stirred at room temperature overnight. The mixture was partitioned with saturated aqueous NaHCC>3, and extracted with CH2CI2 (20 mL x 2). The combined organic layers were washed with brine, dried (Na2S04) and concentrated. The residue was purified by column chromatography (1 % methanol in dichloromethane) to give the desired compound as a yellow oil (2.5 g, 80%): LCMS: RT 2.40min; m/z 334.1 [M+H]\

(d) 4-(3-Oxa-8-azabicyclo[3.2. 1 ]octane-8-carbonyl) -2-ethoxybenzoic acid 172

To a solution of ethyl-4-(3-oxa-8-azabicyclo-[3.2.1]-octane-8-carbonyl)-2-ethoxybenzoate 171 (2.4 g, 7.2 mmol) in a mixture of THF (20 mL), methanol (2 mL) and water (2 mL) was added LiOH H₂0 (1 .5 g, 36 mmol). The resulting mixture was stirred at room temperature for 24 h. The solvent was removed, and the residue obtained diluted with water (20 mL). The pH of the aqueous mixture was adjusted to 6 by addition of a 2 M aqueous HCI solution. The mixture was extracted with CH2CI2 (20 mL x 3) and the combined organic layers washed with brine(10 mL x 2), dried ( a2S0₄) and concentrated to give the desired compound as a yellow oil (1.7 g, 79%): ¹H NMR (400 MHz, A-MeOD) δ 7.83 (d, J = 7.8 Hz, 1 H), 7.19 (d, J = 1.0 Hz, 1 H), 7.10 (dd, J = 7.8, 1.3 Hz, 1 H), 4.65 (br s, 1 H), 4.20 (q, J = 7.0 Hz, 2H), 3.97 (br s, 1 H), 3.82 (d, J = 10.8 Hz, 1 H), 3.72 (d, J = 1 1 .0 Hz, 2H), 3.59 (d, J = 10.9 Hz, 1 H), 2.13 - 1.94 (m, 4H), 1.45 (t, J = 7.0 Hz, 3H).LCMS: RT 1 .20 min; m/z 306.1 [M+H]\

(xxxi 4-(3-Oxa-8-azabicvclo[3.2.1loctane-8-carbonyl)benzoic acid I74

(a) Methyl 4-(3-oxa-8-azabicyclo[3.2. 1 ]octane-8-carbonyl)benzoate 173

To a solution of 4-(methoxycarbonyl)benzoic acid (664 mg, 3.7 mmol) in CH2C (20 mL) was added, 3-oxa-8-azabicyclo[3.2.1 ]octane hydrochloride (500 mg, 3.4 mmol), DIPEA (865.9 mg, 6.7 mmol), HOBt (45 mg, 0.3 mmol) and EDCI (771.4 mg, 4.0 mmol). The resulting mixture was stirred at room temperature overnight. The mixture was partitioned against with aqueous NaHCOs (20 mL x 2) and the aqueous layer extracted with CH2CI2 (5 mL x 2). The combined organic layers were washed with brine (20 mL x 2), dried ( a2S0₄) and concentrated. The crude residue obtained was purified by column chromatography (1 % methanol in CH2CI2) to give the desired compound as a white solid (786.9 mg, 85%): LCMS: RT 0.61 min; m/z 276.1 [M+H]\ (b) 4-(3-Oxa-8-azabicyclo[3.2.1 ]octane-8-carbonyl)benzoic acid 174

To a solution of methyl 4-(3-oxa-8-azabicyclo[3.2.1]octane-8-carbonyl)benzoate I73 (770 mg, 2.8 mmol) in a mixture of THF (20 mL), methanol (2 mL) and water (2 mL) was added LiOH H₂0 (587.8 mg, 14.0 mmol). The resulting mixture was stirred at room temperature overnight, then the solvent removed and the residue obtained diluted with water (20 mL). The pH of the aqueous solution was adjusted to 6 by addition of a 2 M aqueous HCI solution. The aqueous layer was extracted with CH2CI2 (20 mL x 3) and the combined organic layers washed with brine (20 mL x 2), dried ( a2SC>4) and concentrated to give the desired compound (460 mg, 63%) as a white solid: LCMS: RT 0.83 min; m/z 262.1 [M+H]\ xxxii) 2-Methoxv-4-(morpholine-4-carbonyl)benzoic acid I78

(a) (4-Bromo-3-hydroxyphenyl)(morpholino)methanone 175

To a solution of 4-bromo-3-hydroxybenzoic acid (6.0 g, 27.6 mmol) in THF (300 mL) was added HATU (12.6 g, 33.2 mmol), DIPEA (4.3 g, 33.2 mmol) and morpholine (3.6 g, 44.2 mmol). The solution was stirred at room temperature overnight then poured into a solution of 1 M aqueous HCI (400 mL). The aqueous layer was extracted with dichloromethane, washed with water and brine, dried (Na2SO.i). concentrated and the residue purified by column chromatography (0-5% methanol/dichloromethane) to give the desired compound (5.9 g, 75%) as a white solid.

(b) (4-Bromo-3-methoxyphenyl)(morpholino)methanone 176

To a solution of (4-bromo-3-hydroxyphenyl)(morpholino)methanone 175 (8.6 g, 30.0 mmol) in DMF (120 mL) was added iodomethane (5.1 g, 36.1 mmol) and K₂C0₃ (6.2 g, 45.1 mmol). The solution was stirred at room temperature overnight then poured into water (600 ml_). The aqueous layer was extracted with dichloromethane, the combined organic layers were washed with water and brine, dried (Na^-SC ), concentrated and the residue was purified by column chromatography (0-2% methanol in dichloromethane) to give the desired compound (7.5 g, 83%) as a white solid.

(c) Methyl 2-methoxy-4-(morpholine-4-carbonyl)benzoate 177

To a solution of (4-bromo-3-methoxyphenyl)(morpholino)methanone 176 (300 mg, 1.0 mmol) in methanol (10 mL) was added triethylamine (222 mg, 2.2 mmol) and Pd(dppf)C (37 mg, 0.05 mmol). The mixture was heated at reflux under a CO atmosphere overnight. The reaction mixture was concentrated and the residue diluted with water (50 mL) and extracted with EtOAc (50 ml_x2). The organic layer was washed with brine, dried (Na2S0₄), concentrated and the residue was purified by column chromatography (0-5% methanol in dichloromethane) to give the desired compound (200 mg, 80%) as a red oil. LCMS: RT 0.97 min, m/z 280.1 [M+H]⁺

(d) 2-Methoxy-4-(morpholine-4-carbonyl)benzoic acid 178

To a solution of methyl 2-methoxy-4-(morpholine-4-carbonyl)benzoate 177 (200 mg, 0.7 mmol) in a mixture of methanol (0.1 mL), THF (1 mL) and water (0.1 mL) was added UOH.H2O (33 mg, 0.8 mmol). The mixture was stirred at room temperature overnight then poured into water (20 mL) and acidified to pH 6 by addition of a 1 M aqueous HCI solution. The mixture was extracted with dichloromethane (10 mL x 6) and the combined organic layers washed with brine, dried (NazSOn) and concentrated. The residue obtained was purified by column chromatography (0-10% methanol in dichloromethane) to give the desired compound (152 mg, 80%) as a white solid. LCMS: RT 0.41 min, m/z 266.1 [M+H]'

(xxxiii) 6-(3-Oxa-8-azabicvcloi3.2.1 loctane-8-carbonyl)-2-ethoxynicoiinic acid I82

(a) 2-Chloro-6-ethoxypyridine 179

To a solution of 2,6-dichloropyridine (5.0 g, 33.8 mmol) in EtOH (50 mL) was added EtONa (9.2 g, 0.14 mol). The mixture was stirred at 60 °C for 24 h. The solvent was removed and the residue was dissolved in water (100 mL). The aqueous layer was acidified to pH 7 with 2 M aqueous HCI then extracted with DCM (100 mL x 2). The combined organic layers were washed with water (100 mL) and brine (100 mL), dried (Na2S0₄) and concentrated to give the title compound as yellow oil (4.2 g, 79%). LCMS: RT 2.64 min; m/z 158.1 [M+H] '.

(b) 6-Chloro-2-ethoxynicotinaldehyde 180

To a solution of 2-chloro-6-ethoxypyridine 179 (2.0 g, 12.7 mmol) in THF (40 mL) at -78 °C was added i-BuLi (1.6 M in pentane, 8.8 mL, 14.0 mmol) dropwise under N2. After stirring at the same temperature for 1 h, DMF (2.8 g, 38.1 mmol) was added dropwise and the reaction mixture was stirred at -78 °C for 30 min then warmed to room temperature and stirred for 30 min. The reaction mixture was quenched with 2M aqueous HCI (5 mL) and extracted with EtOAc (50 mL x 2). The combined organic layers were washed with water (50 mL x 3) and brine (50 mL), dried (Na2S0₄) and concentrated to give the title compound as yellow oil (2.0 g, 87%): ¹ H NMR (400 MHz, DMSO-d₆) δ 10.17 (s, 1 H), 8.09 (d, J = 8.0 Hz, 1 H), 7.23 (d, J = 8.0 Hz, 1 H), 4.44 (m, 2H), 1 .38 (m, 3H). LCMS: RT 2.64 min; m/z 186 [M+H] '; 218.1

[M+MeOH+H]\

(c) 6-(3-oxa-8-azabicyclo[3.2. 1]octane-8-carbonyl)-2-ethoxynicotinaldehyde 181

To a solution of 3-oxa-8-azabicyclo[3.2.1 ]octane hydrochloride (1 .5 g, 10.3 mmol) in toluene (80 mL) was added Et₃N (2.6 g, 25.8 mmol). Xantphos (0.2 g, 0.34 mmol) and Pd(OAc)₂ (40 mg, 0.17 mmol). The mixture was degassed three times under N2 followed by addition of 6- chloro-2-ethoxynicotinaldehyde I80 (1.6 g, 8.6 mmol). The mixture was degassed three times under N2 and then three times under CO. The mixture was stirred at 90 C overnight. Water (80 mL) was added and the mixture was extracted with EtOAc (80 mL x 2). The combined organic layers were washed with water (80 mL), brine (80 mL), dried (Na₂S0₄) and concentrated. The residue was purified by column chromatography (100% petroleum ether to 50% EtOAc in petroleum ether) to give the title compound as a yellow solid (1.1 g, 44%): LCMS: RT 2.22 min; m/z 291 .1 [M+H]⁺.

(d) 6-(3-Oxa-8-azabicyclo[3.2.1]octane-8-carbonyl)-2-ethoxynicotinic acid 182

To a mixture of 6-(3-oxa-8-azabicyclo[3.2.1]octane-8-carbonyl)-2-ethoxynicotinaldehyde 181 (100 mg, 0.34 mmol) in a mixture of t-BuOH (5 mL) and 2-methylbut-2-ene (2 mL) was added a solution of NaH₂P0₄.2H₂0 (376 mg, 2.4 mmol) and NaCI0₂ (300 mg, 3.3 mmol) in water (5 mL). The resulting mixture was stirred at room temperature for 2 h. The solvent was removed under reduced pressure. The residue obtained was dissolved in water and the aqueous layer was acidified to pH 5 with 2 M aqueous HCI and extracted with EtOAc (20 mL x 4). The combined organic layers were dried (Na₂S0₄) and concentrated to give the title compound as a white solid (80 mg, 76%): LCMS: RT 1 .74 min; m/z 307.1 [M+H]'.

(xxxiv) 2-Ethoxv-4-(5-methyl-1.3.4-oxadiazol-2-vl)benzoic acid I85

I84 I85

(a) Ethyl 4-(2-acetylhydrazinecarbonyl)-2-ethoxybenzoate 183

To a solution of 3-ethoxy-4-(ethoxycarbonyl)benzoic acid 170 (500 mg, 2.1 mmol) in CH₂CI₂ (20 mL) was added DIPEA (960 mg, 7.4 mmol), HOBt (30 mg, 0.2 mmol), EDCI (800 mg, 4.2 mmol) and acetohydrazide (156 mg, 2.1 mmol). The mixture was stirred at room temperature overnight then diluted with CH₂CI₂ (50 mL). The organic layer was washed with water (50 mL) and brine (50 mL), dried (Na₂S0₄) and concentrated. The residue was purified by column chromatography (0- 4% v/v MeOH in CH₂CI₂) to give the title compound as an off- white solid (350 mg, 57%). LCMS: RT 4.79 min; m/z 294.9 [M+H]^*. (b) Ethyl 2-ethoxy-4-(5-methyl- 1 , 3,4-oxadiazol-2-yl)benzoate 184

A mixture of ethyl 4-(2-acetylhyd razi neca rbonyl )-2-ethoxybenzoate 183 (330 mg, 1.1 mmol) in POCI3 (3 mL) was heated at reflux for 1.5 h. The mixture was poured into ice-water (20 mL) and the aqueous layer extracted with CH2CI2 (20 mL x 2). The combined organic layers were washed with a saturated aqueous NaHCO.3 solution (40 mL x 3), brine (40 mL), dried (NazSC ) and concentrated to give the crude product as a yellow solid (290 mg, 94%). The crude product was used for the next step without purification.¹H NMR (400 MHz, CDCI3) δ 7.86 (d, J = 8.4 Hz, 1 H), 7.64 (d, J = 1 .2 Hz, 1 H), 7.57 (m, 1 H), 4.41 -4.35 (m, 2H), 4.23-4.17 (m, 2H), 2.63 (s, 3H), 1.49 (m, 3H), 1 .39 (m, 3H). LCMS: RT 5.40 min; m/z 277.0 [M+H]\

(c) 2-Ethoxy-4-(5-methyl-1,3,4-oxadiazol-2-yl)benzoic acid 185

To a solution of ethyl 2-ethoxy-4-(5-methyl-1 ,3,4-oxadiazol-2-yl)benzoate I84 (270 mg, 1.0 mmol) in MeOH (10 mL) was added a solution of NaOH (193 mg, 4.8 mmol) in water (2 mL). The resulting mixture was stirred at room temperature overnight. The solvent was removed and the residue was suspended in water (5 mL). The pH of the aqueous solution was adjusted to pH 4-5 by addition of 1 M aqueous HCI solution. The solid which precipitated was collected by filtration, washed with water (5 mL) and dried to give the title compound as a yellow solid (1 10 mg, 45%). LCMS: RT 4.77 min, m/z 249.0 [M+H]⁺

(xxxy) 3-(Methylcarbamoyl)benzoic acid I87

B6 I87

(a) Methyl 3-(methylcarbamoyl)benzoate 186

To a solution of isophthalic acid (2.0 g, 12 mmol) in DCM (40mL) with a catalytic amount of DMF (5 drops) at 0 C under 2 atmosphere was added oxalyl chloride (3.81 g, 30 mmol) dropwise. The mixture was allowed to warm to room temperature and stirred for 2 h. The mixture was concentrated under vacuum and the residue was dissolved in DCM (20 mL). 2 M methylamine in tetrahydrofuran solution (9 mL, 18 mmol) was then added dropwise and the reaction stirred at room temperature overnight. The reaction was quenched with MeOH (20 mL), stirred for 30 min then the solvent removed under vacuum. The residue was dissolved in DCM (20 mL) and water (10 mL) and the aqueous layer extracted with DCM (3 x 5 mL). The combined organic extracts were washed with saturated aqueous NaHCOa (3 x 10 mL) and brine (3 x 10 mL), dried ( a2S0₄) and concentrated. The crude product was purified by silica gel chromatography (ethyl acetate/petroleum ether = 1/3 v/v) to give the title compound as a white solid (155 mg, 7%):¹ H NMR (400 MHz, MeOD) δ 8.66 (s, 1 H), 8.42 (s, 1 H), 8.10-8.07 (m, 2H), 7.62 (m, 1 H), 3.88 (s, 3H), 2.80 (m, 3H); LCMS RT 0.91 min; m/z 194.1 [M+H]\

(b) 3-(Methylcarbamoyl)benzoic acid 187

To a solution of methyl 3-(methylcarbamoyl)benzoate 186 (150 mg, 0.78 mmol) in a mixture of THF (4 mL), MeOH (2 mL) and water (1 mL) was added lithium hydroxide monohydrate (163 mg, 3.88 mmol). The resulting mixture was stirred at room temperature overnight. The organic solvent was removed under vacuum and the aqueous acidified to a pH of 1-2 by addition of 3M aqueous HCI solution. The precipitate which formed was collected by filtration, and dried to give the title compound as a white solid (1 15 mg, 82%). LCMS RT 1 .74 min; m/z 180.1 [M+H]\

(xxxvi 4-(3-Oxa-8-azabicvclo[3.2.1loctane-8-carbonyl)-2-chlorobenzoic acid I89

(a) 3-Oxa-8-azabicyclo[3.2.1]octan-8-yl(3-chloro-4-methylphenyl)methanone 188

To a solution of 3-chloro-4-methylbenzoic acid (2.26 g, 13.24 mmol) in DCM (20 mL) was added DIPEA (4.7 g, 36.10 mmol), EDCI (3.5 q. 18.05 mmol), HOBt (190 mg, 1 .40 mmol) and 3-oxa-8-azabicyclo[3.2.1]octane hydrochloride (1 .8g, 12.03 mmol). The resulting mixture was stirred at room temperature overnight. The mixture was diluted with water (10 mL) and partitioned with DCM (10 mL). The aqueous layer was extracted with DCM (3 x 5 mL) and the combined organic extracts washed with saturated aqueous NaHC03 (3 x 5 mL) and brine (3 x 5 mL), dried (Na?S04), filtered and concentrated. The crude product was purified by silica gel chromatography (ethyl acetate /petroleum ether = 1/10) to give the title compound (3.0 g, 92%) as a white solid:¹H NMR (400 MHz, MeOD) δ 7.50 (d, J = 1 .6 Hz, 1 H), 7.40 (d, J = 7.6 Hz, 1 H), 7.33 (m, 1 H), 4.61 -4.56 (m, 1 H), 3.99(s, 1 H), 3.78-3.59 (m, 4H), 2.41 (s, 3H), 2.00-1 .99 (m, 4H); LCMS RT 2.51 min; m/z 266.1 [M+H] ' (b) 4-(3-Oxa-8-azabicyclo[3.2. 1]octane-8-carbonyl)-2-chlorobenzoic acid 189 To a solution of 3-oxa-8-azabicyclo[3.2.1 ]octan-8-yl(3-chloro-4-methylphenyl)methanone I88 (500 mg, 1 .88 mmol) in a mixture of pyridine (5 mL) and water (15 mL) was added KMn0₄ (1 .78 g, 1 1 .28 mmol) in portions. The resulting mixture was heated to 50 °C and stirred 48 h. The mixture was filtered and the filtrate was extracted with ethyl acetate (3 x 5 mL). The pH of the aqueous phase was adjusted pH 1 -2 by addition of concentrated HCI, and then extracted with DCM (8 x 5 mL). The combined organic extracts were dried (Na2S0₄), filtered and concentrated to give the title compound (340 mg, 61 %) as a white solid. LCMS RT 0.88 min; m/z 296.1 [M+H]\

(xxxvii) 2-Ethoxv-5-methyl-4-(morpholine-4-carbonvl)benzoic acid I96

I94 [95 ,96

(a) Methyl 2-hydroxy-4-methylbenzoate 190

To a solution of 2-hydroxy-4-methylbenzoic acid (4.0 g, 26.2 mmol) in methanol (40 mL) was added SOC (4.6 g, 39.4 mmol). The mixture was stirred at 50 C for 1 d. The mixture was concentrated to give the crude product which was dissolved in CH2CI2 (50 mL) and the mixture was washed with water (20 mL) and saturated aqueous NaHCOa solution (20 mL x 3). The organic layer was dried a2S0₄) and concentrated to give the title compound as a brown oil (4.2 g, 96%): LCMS: RT 2.74 min; m/z 167.1 [M+H]\

(b) Methyl 5-bromo-2-hydroxy-4-methylbenzoate 191

To a solution of methyl 2-hydroxy-4-methylbenzoate I90 (2.2 g, 13.24 mmol) in chloroform (20 mL) was added Br2 (2.09 g, 13.24 mmol) dropwise. The mixture was stirred at 0 C for 1 h. The reaction was quenched by addition of sodium sulfite (30 mL, 1 M aqueous solution) and the aqueous layer extracted with CH2CI2 (30 mL x 3). The organic layer was dried

( a2S0₄) and concentrated to give the title compound as yellow oil (3.1 g, 96%): LCMS: RT 3.20 min; m/z 245.0 [M+H]'. (c) Methyl 5-bromo-2-ethoxy-4-methylbenzoate 192

To a solution of methyl 5-bromo-2-hydroxy-4-methylbenzoate 191 (3.1 g, 12.7 mmol) in DMSO (10 mL) at 40 C was added K₂C0₃ (5.26 g, 38.1 mmol) and ethyl bromide (2.07 g, 14.05 mmol) dropwise. The mixture was stirred at 40 °C for 2 h. Further ethyl bromide (2.07 g, 14.05 mmol) was then added dropwise and the resulting mixture was stirred at 40 C for 8 h. The reaction was quenched with water (20 mL) and extracted with CH2CI2 (30 mL x 3). The organic layer was washed with water (20 mL x 10), dried ( a2S0₄ and concentrated to give the title compound as brown oil (2.5 g, 73%): LCMS: RT 2.86 min; m/z 273.0 [M+H] '. (d) 2-Bromo-5-ethoxy-4-(methoxycarbonyl)benzoic acid 193

To a solution of methyl 5-bromo-2-ethoxy-4-methylbenzoate 192 (2.5 g, 9.19 mmol) in pyridine (5 mL) and water (15 mL) was added KMn0₄ (4.3 g, 27.57 mmol). The mixture was stirred at 50 °C for 24 hours, then at room temperature for 2 days. The mixture was filtered through celite, the precipitate was resuspended in 70 C hot water and filtered again. The aqueous solution was washed with EtOAc (30 mL), then acidified with 2M aqueous HCI solution. The mixture was extracted with CH₂CI₂ (50 mL x 5) and the combined organic layers dried (Na2S0₄) and concentrated to give the title compound as a white solid (610 mg, 23%). LCMS: RT 2.26 min; m/z 303.1 [M+H]'. (e) Methyl 5-bromo-2-ethoxy-4-(morpholine-4-carbonyl)benzoate 194

To a solution of 2-bromo-5-ethoxy-4-(methoxycarbonyl)benzoic acid 193 (600 mg, 1.98 mmol) in CH2CI2 (10 mL) at 0 C was added oxalyl chloride (377 mg, 2.97 mmol) and DMF (0.01 mL). The mixture was stirred at 0 °C for 3 h then morpholine (344.9 mg, 3.96 mmol) was added to the mixture, followed by triethylamine (280 mg, 2.77 mmol). The resulting mixture was stirred at room temperature overnight. The reaction was poured into water (30 mL) and the aqueous layer extracted with CH2CI₂ (50 mL x 3). The organic layer was dried (Na2SO.t) and concentrated and the residue was purified by column chromatography (CH2CI2: methanol=100:1 v/v) to give the title compound as red oil (790 mg, 95%): LCMS: RT 2.30 min; m/z 372.0 [M+H] '.

(f) Methyl 2-ethoxy-5-methyl-4-(morpholine-4-carbonyl)benzoate 195

To a solution of methyl 5-bromo-2-ethoxy-4-(morpholine-4-carbonyl)benzoate 194 (360 mg, 0.97 mmol) in DMF (10 mL) was added K2CO3 (268.1 mg, 1 .94 mmol), methyl boronic acid (1 16 mg, 1 .94 mmol) and Pd(PPh3)₄ (56 mg, 0.05 mmol). The resulting mixture was heated at 100 °C under N2 for 1 day. Water (20 mL) was added to the mixture, and the aqueous extracted with CH2C (20 mL x 3). The organic layer was washed with brine (20 mL), dried (Na2S04) and concentrated. The residue was purified by column chromatography (CH2CI2: methanol=100:1 v/v) to give the title compound as yellow oil (100 mg, purity about 80%). LCMS: RT 2.08 min; m/z 308.1 [M+H]\ (g) 2-Ethoxy-5-methyl-4-(morpholine-4-carbonyl)benzoic acid 196

To a solution of methyl 2-ethoxy-5-methyl-4-(morpholine-4-carbonyl)benzoate 195 (100 mg, 0.32 mmol) in a mixture of THF (10 mL), methanol (1 mL) and water (0.1 mL) was added LiOH H2O (41 mg, 0.96 mmol). The resulting mixture was stirred at room temperature for 1 day. The reaction mixture was concentrated and the residue diluted with water (20 mL). The pH of the aqueous mixture was adjusted to 6 by addition of 1 M aqueous HCI solution and the aqueous layer extracted with CH2CI2 (20 mL x 3). The combined organic layers were dried ( a2S0₄) and concentrated and the crude product purified by preparative TLC (CH2CI2: methanol=10:1 v/v) to give the title compound as a white solid (20 mg, 45%). LCMS: RT 0.85 min; m/z 294.1 [M+H]\

(xxxviii) 6-(3-Oxa-8-azabicvclo[3. . ?loctane-8-carbonvl)-4-ethoxynicotinic acid 1102

(a) Ethyl 4-ethoxypicolinate 197

A solution of 4-chloropicolinonitrile (5.5 g, 39.7 mmol) in a saturated solution of HCI in EtOAc (80 mL) was stirred at 80 C for 2 days. The solvent was removed under reduced pressure. Saturated aqueous NaHCC solution (200 mL) was added and the aqueous layer was extracted with CH2CI2 (3 x 200 mL). The combined organic extracts were dried ( a2S0₄) and concentrated to give the title compound as a white solid (2.6 g, 34%): ¹ H NMR (400 MHz, CDCI3) δ 8.52 (d, J = 5.6 Hz, 1 H), 7.63 (d, J = 2.4 Hz, 1 H). 6.93-6.91 (m, 1 H), 4.80-4.43 (m, 2H), 4.16-4.1 1 (m, 2H), 1.46-1 .41 (m, 6H); LCMS: RT 1.35 min; m/z 196.1 [M+H]⁺. (b) Ethyl 5-bromo-4-ethoxypicolinate 198

To a solution of ethyl 4-ethoxypicolinate 197 (1 .6 g, 8.2 mmol) in concentrated H2SO4 (80 mL) was added /V-bromosuccinimide (2.7 g, 14.8 mmol). The reaction was stirred at room temperature overnight and then quenched by addition of a saturated aqueous NaHCO.3 solution (150 mL). The aqueous layer was extracted with CH2CI2 (3 x 130 mL) and the combined organic layers were dried ( a2S0₄) and concentrated to give the title compound as a yellow solid (2.0 g, 91 %):¹H NMR (400 MHz, CDC ) «5 8.67 (s, 1 H), 7.63 (s, 1 H), 4.94- 4.40 (m, 2H), 4.29-4.23 (m, 2H), 1 .54-1 .51 (m, 3H),1 .46-1.42 (m, 3H). LCMS: RT 2.59 min; m/z 274.0 [M+H]' for ⁷⁹Br, 276.0 [M+H]⁺ for ⁸¹ Br.

(c) 5-Bromo-4-ethoxypicolinic acid 199

To a solution of ethyl 5-bromo-4-ethoxypicolinate 198 (1.6 g, 6.0 mmol) in a mixture of THF (20 mL), CH3OH (2 mL) and H₂0 (0.2 mL) was added ϋΟΗΉ₂0 (1 .0 g, 24.0 mmol). The reaction was stirred at room temperature for 2 d. The solvent was removed under reduced pressure and the residue was dissolved in water (10 mL) and acidified to pH 3 with 1 M aqueous HCI. The aqueous phase was extracted with CH2CI2 (4 x 50 mL) and the combined organic extracts were dried ( a2S0₄) and concentrated to give the title compound as a yellow solid (800mg, 54%). LCMS: RT 0.86 min; m/z 246.0 [M+H]' for ⁷⁹Br, 248.0 [M+H]' for ⁸¹ Br.

(d) 3-Oxa-8-azabicyclo[3.2.1]octan-8-yl(5-bromo-4-ethoxypyridin-2yl)methanone 1100 To a solution of 5-bromo-4-ethoxypicolinic acid I99 (300 mg, 1.2 mmol) and 3-oxa-8- azabicyclo[3.2.1]octane(153 mg, 1.0 mmol) hydrochloride salt in CH2CI2 (5 mL) were added DIPEA (0.72 mL, 4.1 mmol), EDCI (393 mg, 2.0 mmol) and HOBt (15 mg, 0.1 mmol).The mixture was stirred at room temperature overnight. The reaction was quenched by addition of saturated aqueous NaHCCb solution (50 mL) and the mixture was extracted with CH2CI2 (3 x 50 mL). The combined organic layers were dried ( a2S0₄) and concentrated and the residue was purified by silica gel chromatography (20% EtOAc in petroleum ether v/v) to give the title compound as a yellow solid (262 mg, 75%). LCMS: RT 5.56 min; m/z 340.9 [M+H] ' for ⁷⁹Br, 342.9 [M+H]' for ⁸¹ Br.

(e) Methyl 6-(3-oxa-8-azabicyclo[3.2. 1]octane-8-carbonyl)-4-ethoxynicotinate 1101

To a solution of 3-oxa-8-azabicyclo[3.2.1 ]octan-8-yl(5-bromo-4-ethoxypyridin-2- yl)methanone 1100 (650 mg, 1.9 mmol) in MeOH (50 mL) was added Et₃N (577 mg, 5.7 mmol) and PdC (dppf) (73 mg, 0.05 mmol). The reaction was stirred under a CO

atmosphere and heated at reflux for 2 days. The solvent was removed under reduced pressure and the residue was taken up in water (100 mL) and extracted with CH2CI2 (3 x 100 mL). The combined organic layers were dried over ( a2S0₄) and concentrated. The residue was purified by silica gel chromatography (33% EtOAc in petroleum ether v/v) to give the title compound as a brown solid (400 mg, 66%). LCMS: RT 4.94 min; m/z 320.8 [M+H]\

(f) 6- (3- Oxa -8-azabicyclo[3.2.1]octane-8-carbonyl)-4-ethoxynicotinic acid 1102

To a solution of methyl 6-(3-oxa-8-azabicyclo[3.2.1 ]octane-8-carbonyl)-4-ethoxynicotinate 1101 (350 mg, 1 .1 mmol) in CH₃OH (20 mL) was added NaOH (2.2 mL of a 1 M aqueous solution) and the mixture stirred at room temperature overnight. The solvent was removed under reduced pressure and the residue was taken up in water (15 mL). The aqueous was acidified to pH 4 by addition of 1 M aqueous HCI. The aqueous phase was extracted with CH2CI2 (4 x 60 mL) and the combined extracts were dried ( a2S0₄) and concentrated to give the title compound as a yellow solid 1102 (320 mg, 95%). LCMS: RT 0.61 min; m/z 307.1 [M+H]\ (xxxix) 2-Ethoxv-3-methyl-4-(morpholine-4-carbonvl)benzoic acid 1109

1107 1108 1109

(a) Methyl 3-hydroxy-2-methylbenzoate 1103

To a solution of 3-hydroxy-2-methylbenzoic acid (10.0 g, 65.7 mmol) in MeOH (100 mL) was added SOCI2 (15.6 g, 131.5 mmol) slowly and the mixture was stirred at 50 °C overnight. The reaction mixture was concentrated and the residue dissolved in DCM (100 mL). The organic solution was washed with saturated aqueous NaHCOa solution, dried ( a2S0₄) and concentrated to give the title product (10.7 g, 98%) as white solid. LCMS: RT 1.98 min; m/z 167.1 [M+H]\

(b) Methyl 4-bromo-3-hydroxy-2-methylbenzoate 1104

To a solution of iert-butylamine (2.0 g, 27.1 mmol) in DCM (180 mL) at -70 ' C was added a solution of Br₂ (4.2 g, 27.1 mmol) in DCM (10 mL) dropwise and the mixture was stirred at - 70 C for 1 h. A solution of methyl 3-hydroxy-2-methylbenzoate 1103 (4.5 g, 27.1 mmol) in DCM (10 mL) was then added dropwise and the resulting mixture allowed to warm to room temperature and stirred overnight. The reaction was quenched by addition of water (30 mL) and the aqueous layer extracted with DCM (3 x 50 mL). The combined organic layers were washed with brine, dried (IS^SC ) and concentrated. The crude residue obtained was purified by silica gel chromatography (2% EtOAc in petroleum ether v/v) to give the title product (2.2 g, 33%) as a white solid. LCMS: T 2.46 min; m/z 245.0 [M+H]⁺ for ⁷⁹Br, 247.0 [M+H]' for ⁸¹ Br. H NMR (400 MHz, d₆-DMSO) δ 9.38 (s, 1 H), 7.46 (d, J = 8.4 Hz, 1 H), 7.17 (d, J = 8.4 Hz, 1 H). 3.80 (s, 3H), 2.38 (s, 3H).

(c) Methyl 4-bromo-3-ethoxy-2-methylbenzoate 1105

To a solution of methyl 4-bromo-3-hydroxy-2-methylbenzoate 1104 (2.1 g, 8.6 mmol) in DMSO (10 mL) was added K₂C0₃ (3.5 g, 25.8 mmol) and ethyl bromide (1 .4 g, 12.9 mmol). The reaction was stirred at 40 C overnight. Water (30 mL) was added and the aqueous extracted with DCM (3 x 20 mL). The organic layer was washed with water (10 x 30 mL), dried (Na2SO.i), and concentrated to give the title product (2.2 g, 95%) as a yellow solid. LCMS: RT 3.08 min; m/z 273.0 [M+H]' for ⁷⁹Br, 275.0 [M+H]' for ⁸¹Br.

(d) 4-Bromo-3-ethoxy-2-methylbenzoic acid 1106

To a solution of methyl 4-bromo-3-ethoxy-2-methylbenzoate 1105 (2.1 g, 10.0 mmol) in a mixture of MeOH (10 mL) and water (0.2 mL) was added NaOH (0.4 g, 20.0 mmol). The reaction was stirred at room temperature overnight and then the solvent was removed under reduced pressure. The residue obtained was dissolved in water (20 mL) and acidified with 1 M aqueous HCI to pH 2. The aqueous layer was extracted with DCM (4 x 20 mL) and the combined organic layers dried ( a2S0₄) and concentrated to give the title product (2.4 g, 92%) as a white solid. LCMS: RT 2.75 min; m/z 281 .0 [M+Na]' for ⁹Br., 283.0 [M+Na]' for ⁸ Br.

(e) (4-Bromo-3-ethoxy-2-methylphenyl)(morpholino)methanone 1107

To a solution of 4-bromo-3-ethoxy-2-methylbenzoic acid 1106 (2.4 g, 9.3 mmol) in DCM (500 mL) at 0 °C was added oxalyl chloride (3.5 g, 27.9 mmol) and DMF (0.2 mL). The reaction was stirred for 3 h then morpholine (1.6 g, 18.6 mmol) and triethylamine (4.1 g, 40.9 mmol) were added and stirring was continued overnight at 0 °C. Water (100 mL) was added and the mixture was extracted with DCM (2 x 100 mL). The combined organic layers were washed with brine, dried ( a2S0₄) and concentrated. The residue was purified by silica gel chromatography (1 % MeOH in DCM v/v) to give the title compound (720 mg, 24%) as a white solid which was used without further purification. LCMS: RT 2.48 min; m/z 328.1

[M+H]⁺ for ⁷⁹Br, 330.1 [M+H]' [M+H]⁺ for ⁸¹Br.

(f) Methyl 2-ethoxy-3-methyl-4-(morpholine-4-carbonyl)benzoate 1108

To a solution of (4-bromo-3-ethoxy-2-methylphenyl)(morpholino)methanone 1107 (720 mg, 2.2 mmol) in MeOH (10 mL) was added PdC (dppf) (81 mg, 0.1 1 mmol) and triethylamine (489 mg, 4.8 mmol). The reaction was stirred under a CO atmosphere and heated at reflux overnight. The mixture was concentrated and the residue was dissolved in DCM (20 mL), washed with water and brine, dried ( a2S0₄) and concentrated. The residue obtained was purified by silica gel chromatography (1 % MeOH in DCM v/v) to give the title product (670 mg, 80% purity by HPLC) as yellow oil. LCMS: RT 2.02 min; m/z 308.2 [M+H]'.

(g) 2-Ethoxy-3-methyl-4-(morpholine-4-carbonyl)benzoic acid 1109

To a solution of methyl 2-ethoxy-3-methyl-4-(morpholine-4-carbonyl)benzoate 1108 (670 mg, 2.2 mmol) in a mixture of THF (1 mL), MeOH (10 mL) and water (0.1 mL) was added

LiOH'H₂0 (275 mg, 6.5 mmol) The reaction was stirred at room temperature overnight and then concentrated under reduced pressure. The residue was dissolved in water (20 mL) then acidified with 1 M aqueous HCI to pH 2. The aqueous mixture was extracted with DCM (10 mL x 3) and the combined organic layers were dried ( a2S0₄) and concentrated to give the title product (310 mg, 48%) as brown oil. LCMS: RT 0.89 min; m/z 294.1 [M+H]'.

(xl) 2-Fluoro-4-(3-oxa-8-aza-bicvclo[3.2.11octane-8-carbonyl)-benzoic acid 11 1 1

(a) (3-Fluoro-4-methyl-phenyl)-(3-oxa-8-aza-bicyclo[3.2. 1 ]oct-8-yl)-methanone 1110

To a solution of 3-fluoro-4-methylbenzoic acid (2.0 g, 13.0 mmol) and 3-oxa-8- azabicyclo[3.2.1]octane hydrochloride salt (1 .8 g, 1 1.8 mmol) in CH₂CI₂ (10 mL) were added DIPEA (6.3 mL, 35.4 mmol), EDCI (2.72 g, 14.2 mmol) and HOBt (162.1 mg, 1 .2 mmol). The mixture was stirred at room temperature overnight. A saturated aqueous NaHC03 solution (50 mL) was added and the mixture was extracted with CH₂C (3 x 50 mL). The combined organic exctracts were dried (Na₂S0₄) and concentrated. The residue was purified by silica gel chromatography (5% EtOAc in petroleum ether v/v) to give the title compound as a yellow solid (1.7 g, 58%). LCMS: RT 2.30 min; m/z 250.1 [M+H]\ (b) 2-Fluoro-4-(3-oxa-8-aza-bicyclo[3.2.1 ]octane-8-carbonyl)-benzoic acid 1111 To a solution of (3-fluoro-4-methyl-phenyl)-(3-oxa-8-aza-bicyclo[3.2.1 ]oct-8-yl)-methanone 1110 (800 mg, 3.2 mmol) in a mixture of pyridine (6 mL) and H2O (12 mL) was added KMnC (5.1 g, 32 mmol) and the mixture stirred at room temperature for 2 days. The resulting suspension was filtered through Celite and the filtrate washed with CH2C (3 x 50 mL). The remaining aqueous layer was adicified to pH 3 by addition of 2 M aqueous HCI, and extracted with CH2CI2 (4 x 60 mL). The combined organic layers were dried a2S0₄) and concentrated to give the title compound 1111 as an off-white solid (500 mg, 56%). LCMS: RT 2.89 min; m/z 280.1 [M+H]^*.

(xli 4-(3-Oxa-8-azabicvclo[3.2.1 loctane-8-carbonyl)-2-ethoxvbenzoic acid 11 15

11 14 1115

(a) Methyl 2-methoxy-4-methylbenzoate 11 12

To a mixture of 2-hydroxy-4-methylbenzoic acid (10.0 g, 65.7 mmol) and K2CO3 (22.7 g,

164.3 mmol) in DMF (200 mL) at room temperature was added methyl iodide (20.5 g, 144.5 mmol) over a period of 10 min. The resulting mixture was stirred at room temperature overnight, then diluted with CH2CI2 (150 mL) and filtered. The filtrate was washed with water (200 mL x 10) and brine (200 mL x 2), dried (Na2SO.t) and concentrated to give the title compound as a yellow liquid (1 1 .0 g, 93%).

(b) 3-Methoxy-4-(methoxycarbonyl)benzoic acid 11 13

To a solution of methyl 2-methoxy-4-methylbenzoate 11 12 (10.0 g, 48.05 mmol) in a mixture of pyridine (30 mL) and water (90 mL) was added KMn0₄ (30.37 g, 192.2 mmol). The resulting mixture was heated at 50 °C for 48 h, then cooled and allowed to stir at room temperature for 24 h. The mixture was filtered and the filter cake washed with hot water. The combined aqueous filtrates were washed with EtOAc (75 mL x 3) and acidified to pH 2 with a 2 M aqueous HCI solution. The mixture was extracted with CH2CI2 (150 mL x 3). The combined organic layers were washed with brine, dried (Na2S04) and concentrated to give the title compound as a white solid (7.0 g, 61 %). LCMS: RT 1.24 min; m/z 21 1 .0 [M+H]\

(c) Methyl 4-(3-oxa-8-azabicyclo[3.2.1]octane-8-carbonyl)-2-methoxybenzoate 1114

To a solution of 3-methoxy-4-(methoxycarbonyl)benzoic acid 11 13 (500 mg, 2.10 mmol) in CH2CI2 (20 mL) was added 3-oxa-8-azabicyclo[3.2.1 ]octane hydrochloride (377 mg, 2.52 mmol), HOBt (426 mg, 3.15mmol), triethylamine (850 mg, 8.4 mmol) and EDCI (602 mg, 3.15 mmol). The resulting mixture was stirred at room temperature overnight. Saturated aqueous NaHCOa solution was added. The organic layer was separated and the aqueous phase was extracted with CH2CI2 (20 mL x 2). The combined organic layers were washed with brine, dried (Na2S0₄) and concentrated. The residue was purified by column

chromatography (5% methanol in dichloromethane v/v) to give the title compound as a white solid (800 mg, 59%). (d) 4-(3-Oxa-8-azabicyclo[3.2.1 ]octane-8-carbonyl)-2-methoxybenzoic acid 11 15

To a solution of methyl 4-(3-oxa-8-azabicyclo[3.2.1]octane-8-carbonyl)-2-methoxybenzoate 11 14(0.9 g, 2.9 mmol) in a mixture of THF (4 mL) and methanol (4 mL) was added aqueous NaOH solution (2 M, 4 mL). The resulting mixture was stirred at room temperature for 14 h. The solvent was removed, and the residue was diluted with water (20 mL). The pH of the aqueous mixture was adjusted to 6 by addition of 2 M aqueous HCI solution. The mixture was extracted with CH2C (20 mL x 3) and the combined organic layers washed with brine (10 mL x 2), dried ( a2S0₄) and concentrated to give the title compound as a white solid (0.7 g, 83%):LCMS: RT 0.55 min; m/z 292.1 [M+H]\ ( -tert-butyl(3-amino-2-hydroxypropyl)(2,3-dihydro-1 H-inden-2- l)carbamate 11 17

(a) (R)-tert-Butyl(2,3-dihydro- 1 H-inden-2-yl)(3-(1 , 3-dioxoisoindolin-2-yl)-2- hydroxypropyl)carbamate 11 16

A mixture of 2,3-dihydro-1 H-inden-2-amine (5.0 g, 24.61 mmol) and (f?)-2-(oxiran-2- ylmethyl)isoindoline-1.3-dione (4.91 g, 36.91 mmol) in EtOH (100 mL) was heated at reflux overnight. After cooling to room temperature, triethyamine (9.94 g, 98.44 mmol) and (Βο Ο (10.74 g, 49.22 mmol) were added and the reaction mixture was stirred at room temperature overnight. The mixture was concentrated and purified by column (EtOAc / Pet. Ether = 1/4, v/v) to give the title compound (5.4 g, 50%) as an oil.

(b) (R)-tert-butyl(3-amino-2-hydroxypropyl)(2,3-dihydro-1H-inden-2-yl)carbamate 11 17 A mixture of (fl)-ie/?-butyl(2,3-dihydro-1 H-inden-2-yl)(3-(1 ,3-dioxoisoindolin-2-yl)-2- hyd roxypropy I )ca rba mate 11 16 (4.4 g, 10. 08 mmol) and NH₂NH₂.H₂0 (80%, 1 .89 g, 30. 24 mmol) in EtOH (100 mL) was heated at reflux for 4 h. After cooling to room temperature, the mixture was filtered and the filtercake was washed with EtOH (50 mL). The filtrate was concentrated and then dissolved in CH2CI2, washed with saturated aqueous solution NaHCOa (80 mL x 3) and brine (20 mL x 2), dried (Na₂S0₄) and concentrated to give the title compound (3.1 g, crude) as a yellow oil, which was used for the next step without further purification. LCMS: RT 2.17 min; m/z 307.2 [M+H]\

(a) Ethyl 2-ethoxy-4-methylbenzoate 11 18

To a mixture of 2-hydroxy-4-methylbenzoic acid (20.0 g, 131.5 mmol) and K₂COa (54.5 g, 394.5 mmol) in DMSO (50 mL) at 40 °C was added bromoethane (21 .5 g, 197.2 mmol) over 30 min. The reaction was stirred at 40 C for 2h. Further bromoethane (21 .5 g, 197.2 mmol) was then added dropwise and the reaction stirred at 40 °C for 8h. DCM (200 mL) was added and the mixture was filtered. The organic layer was separated washed with water (150 mL x 4), dried (Na₂S0₄) and concentrated to give the title product (24.1 g, 88%) as clear oil. LCMS: RT 2.77 min; m/z 209.1 [M+H] ⁺.

(b) 2-(3-ethoxy-4-(ethoxycarbonyl)phenyl)acetic acid 11 19

To a solution of diisopropylamine (1 .46 g, 14.4 mmol) in dry THF (20 ml) at -30 °C was added n-BuLi (2.4 M in hexane, 5.8 mL, 14.4 mmol) slowly under nitrogen atmosphere. The mixture was stirred at -30 °C for 30 min then cooled to -78 °C and HMPA (4.0 g) added. A solution of ethyl 2-ethoxy-4-methylbenzoate 11 18 (2.0 g, 9.6 mmol) in dry THF (5 mL) was then added dropwise and the resultant mixture stirred at -78 C for 2 h. CO2 (g) was then bubbled through the mixture for a further 30 min until the colour disappeared. The reaction was allowed to warm to 10 C then was diluted with water (20 mL) and extracted with ether (20 mL x 2). The aqueous layer was acidified to pH 2 by addition of 10% aqueous H2SO4 and the mixture extracted with DCM (20 mL x 2). The combined DCM layers were washed with water and brine, dried ( a2S0₄) and concentrated to give the title compound (550 mg, 23%) as yellow oil. LCMS: RT 2.36 min; m/z 253.1 [M+H]⁺.

(c) Ethyl 4-(2-(3-oxa-8-azabicyclo[3.2. 1]octan-8-yl)-2-oxoethyl)-2-ethoxybenzoate 1120 A mixture of 2-(3-ethoxy-4-(ethoxycarbonyl)phenyl)acetic acid 11 19 (550 mg, 2.2 mmol), 3- oxa-8 -azabicyclo[3.2.1] octane hydrochloride salt (320 mg, 2.1 mmol), EDCI (500 mg, 2.6 mmol), HOBt (30 mg, 0.22 mmol) and DIEA (710 mg, 5.5 mmol) in DCM (10 mL) was stirred at room temperature for overnight. Water (20 mL) was added and the reaction mixture extracted with DCM (10 mL x 3), the organic layers were combined and washed with saturated aqueous NaHCOa, water and brine, dried a2SC¼) and concentrated to give the title product (710 mg, 93%) as yellow oil. LCMS: RT 2.38 min; m/z 348.2 [M+H]\ (d) 4-(2-(3-oxa-8-azabicyclo[3.2. 1]octan-8-yl)-2-oxoethyl)-2-ethoxybenzoic acid 1121 To a solution of ethyl 4-(2-(3-oxa-8-azabicyclo[3.2.1 ]octan-8-yl)-2-oxoethyl)-2- ethoxybenzoate 1120 (710 mg, 2.0 mmol) in a mixture of THF (5 mL), MeOH (10 mL) and water (5 mL) was added UOH.H2O (429 mg, 10.2 mmol). The reaction was stirred at room temperature overnight then the solvent removed under vacuum. The residue obtained was dissolved in water (10 mL) then acidified to pH 2 by addition of 10% aqueous H2SO4 solution. The resulting mixture was extracted with DCM (10 mL x 3) and the organic layers dried ( a2S0₄) and concentrated to give the title product (540 mg, 85%) as yellow solid. LCMS: RT 1 .20 min; m/z 320.2 [M+H]\ fxliv) (S -fe/i-butyl(3-amino-2-hvdroxypropyl (2.3-dihvdro-1 H-inden-2-yl)carbamate 1123

(a) (S)-ten-butyl(2,3-dihydro-1H-inden-2-yl)(3-(1,3-dioxoisoind

hydroxypropyl)carbamate 1122

A mixture of 2,3-dihydro-1 H-inden-2-amine (5.0 g, 24.61 mmol) and (S)-2-(oxiran-2- ylmethyl)isoindoline-1.3-dione (4.91 g, 36.91 mmol) in EtOH (100 mL) was heated at reflux overnight. After cooling to room temperature, triethylamine (9.94 g, 98.44 mmol) and (Boc)₂0 (10.74 g, 49.22 mmol) were added and the reaction mixture was stirred at room temperature overnight. The mixture was concentrated and purified by silica gel column chromatography (EtOAc / petroleum ether = 1/4 v/v) to give the title compound (5.4 g, 50%) as an oil. H NMR (400 MHz, DMSO-d₆) δ 7.88-7.82 (m, 4H), 7.16-7.09 (m, 4H), 6.28 (d, J = 4.4 Hz, 1 H), 4.51 (br s, 1 H), 4.05-3.97 (m, 1 H), 3.61 -3.38 (m, 3H), 3.32-3.09 (m, 5H), 1.31 (s, 9H).

(b) (S)-tert-butyl(3-amino-2-hydroxypropyl)(2,3-dihydro- 1 H-inden-2-yl)carbamate 1123 A mixture of (S)-ieri-butyl(2.3-dihydro-1 H-inden-2-yl)(3-(1 ,3-dioxoisoindolin-2-yl)-2- hyd roxypropyl )ca rba mate 1122 (5.0 g, 1 1 .46 mmol) and NH₂NH₂'H₂0 (80%, 2.2 g, 34.37 mmol) in EtOH (100 mL) was heated at reflux for 4 h. After cooling to room temperature, the mixture was filtered and the filter cake was washed with EtOH (50 mL). The filtrate was concentrated and then dissolved in CH2CI2, washed with saturated aqueous solution NaHCOa (80 mL x 3), brine (20 mL x 2), dried (Na₂S0₄) and concentrated to give the title compound (3.4 g, crude) as a yellow oil, which was used for the next step without further purification. LCMS: RT 1 .99 min; m/z 307.2 [M+H]⁺.

Example 1 : 4-ChIoro-iV-(3-((2,3-dihydro-1 H-inden-2-yI);

h droxypropyl)benzamide (1)

A solution of 4-chlorobenzoic acid (0.057 g, 0.36 mmol) in A/,A/-dimethylformamide (2 mL) containing A/,/V-diisopropylethylamine (0.230 mL, 1 .31 mmol), V-ethyl- V-(3- dimethylaminopropyl)carbodiimide hydrochloride (0.095 g, 0.49 mmol) and 1 - hydroxybenzotriazole hydrate (wetted with not less than 20 wt. % water,0.095 g, 0.49 mmol) was stirred at room temperature for 20 minutes. A solution of 1 -amino-3-((2.3-dihydro-1 H- inden-2-yl)amino)propan-2-ol I2 (0.068 g, 0.33 mmol) in DCM (1 mL) was added and stirred at room temperature for 20 hours under a nitrogen atmosphere. Water and ethyl acetate were added. The aqueous phase was extracted with ethyl acetate (2 20 mL) and the combined organic extracts were washed with water (2 20 mL), brine, dried (sodium sulfate), filtered and concentrated in vacuo. The resulting residue was suspended in diethyl ether, sonicated for 10 seconds, filtered and dried to give the desired compound as cream coloured solid (0.090 g, 79%). ¹ H NMR (300 MHz, d,.methanol) δ 7.81 (d, J=8.55 Hz, 2H), 7.47 (d. J=8.55 Hz, 2H), 7.20 - 7.10 (m, 4H), 3.98 - 3.90 (m, 1 H), 3.68-3.59 (m, 1 H), 3.48 (d, J=5.37 Hz, 2H), 3.20 (ddd, J=15.7, 7.15, 4.0 Hz, 2H), 2.84 - 2.76 (m, 3H), 2.73 - 2.66 (m, 1 H). LCMS-D: RT 4.85 min; m/z 345 [M+H]\

Example 2: A -(3-((2,3-Dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)-4-(morpholine-4- carbon l)benzamide (2)

(a) Ιβύ- Butyl (2-hydroxy-3-(4-(morpholine-4-carbonyl)benzamido)propyl)carbamate (A 1) To a solution of 4-(4-morphlinylcarbonyl)benzoic acid I27 (400 mg, 1.70 mmol) in DCM (15 mL) was added Λ/,/V-diisopropylethylamine (1.19 mL, 6.80 mmol), A/-ethyl-/V-(3- dimethylaminopropyl)carbodiimide hydrochloride (489 mg, 2.55 mmol) and 1 - hydroxybenzotriazole hydrate (390 mg, 2.55 mmol) and the mixture was stirred at room temperature for 20 minutes. The solution was cooled to 0 C and A/-Boc-1 ,3-diamino-2- propanol (356 mg, 1 .87 mmol) was added. After stirring at 0°C for 10 minutes, the mixture was stirred at room temperature for 20 hours whilst under a nitrogen atmosphere. Water (15 mL) and DCM (20mL) were added. The aqueous phase was extracted with DCM (2 30 mL) and the combined organic extracts were washed with brine (60 mL), dried (sodium sulfate), filtered and concentrated under reduced pressure to give the desired

compound(450 mg, 65%). LCMS-D: RT 4.60 min; m/z 408 [M+H]\ (b) N-(3-Amino-2-hydroxypropyl)-4-(morpholine-4-carbonyl)benzamide hydrochloride (A2) A solution of fe/f-butyl (2-hydroxy-3-(4-(morpholine-4-carbonyl)benzamido)propyl)carbamate A1 (450 mg, 1 .10 mmol) in 4.0 M hydrochloric acid in 1.4-dioxane (1 .38 mL, 5.52 mmol) and 1 ,4-dioxane (5 mL) was heated at 60 C for 2 hours under a nitrogen atmosphere. The solvent was removed under reduced pressure to give the desired compound (271 mg, 72%).

(c) N -(3-((2,3-Dihydro- 1H-inden-2-yl)amino)-2-hydroxypropyl)-4-(morpholine-4- carbonyl)benzamide (2)

A/-(3-amino-2-hydroxypropyl)-4-(morpholine-4-carbonyl)benzamide hydrochloride A2 (about 200 mg) was dissolved in methanol (6 mL) and applied to an SCX cartridge (5 g). The cartridge was washed with methanol (40 mL) and eluted with 2 M ammonia in methanol (20 mL). The ammonia wash was concentrated to give the free base (163 mg, 0.53 mmol). The material was dissolved in 1 ,2-dichloroethane (10 mL) and THF (5mL) and diisopropylethyl amine (92.4 μί, 0.53 mmol) was added. To the resulting solution, was added 2-indanone (76.5 mg, 0.18 mmol), sodium triacetoxyborohydride (592 g, 2.65 mmol, 95%) and glacial acetic acid (61 L, 1.06 mmol). The mixture was stirred at room temperature for 20 hours under a nitrogen atmosphere. The solution was made basic using 10 M aqueous NaOH, then water (15 mL) and ethyl acetate (15 mL) were added. The aqueous phase was extracted with ethyl acetate (2 20 mL) and the combined organic extracts were washed with water (20 mL), brine (20 mL), dried (sodium sulfate), filtered and concentrated in vacuo.T e material was purified by column chromatography (Si02, 0-20% methanol/DCM) to give the desired compound (77 mg, 34%); ¹ H NMR (300 MHz, CDCb) 7.83 (d, J = 8.4 Hz, 2H), 7.64 (m, 1 H), 7.40 (d, J = 8.4 Hz, 2H), 7.17 (m, 4H), 3.92 (br m, 3H), 3.77 (m, 3H), 3.66 (m, 4H), 3.45 - 3.37 (m, 3H), 3.19 (dd, J = 15.7, 7.2 Hz, 2H), 2.96 - 2.84 (m, 3H), 2.73 (dd, J = 12.2, 7.6 Hz, 1 H). LCMS-D: RT 4.15 min; m/z 424.3 [M+H]\

Example 3: (fl)-/V-(3-{(2,3-Dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)- -methyl-2- (4- trifluoromethyl)phenyl)thiazole-5-carboxamide (3)

A solution of 4-methyl-2-(4-(trifluoromethyl)phenyl)-1 ,3-thiazole-5-carboxylic acid (101 mg, 0.352 mmol) in DCM (4.0 mL) containing A/.A/-diisopropylethylamine (0.184 mL, 1.06 mmol), N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide hydrochloride (88 mg, 0.46 mmol) andl - hydroxybenzotriazole hydrate (70 mg, 0.46 mmol) was stirred for 20 minutes. 1-Amino-3- ((2,3-dihydro-1 H-inden-2-yl)amino)propan-2-ol I4 (65 mg, 0.32 mmol) in DCM (1 mL) was added and the mixture was stirred at room temperature for 24 hours under a nitrogen atmosphere. Water (15 mL) and DCM (15 mL) were added. The aqueous phase was extracted with DCM (2 15 mL) and the combined organic extracts were washed with brine (20 mL), dried (sodium sulfate), filtered and concentrated in vacuo. The residue was dissolved in methanol and applied to an SCX cartridge (1 g). The cartridge was washed with methanol (10 mL) and eluted with 2M ammonia in ethanol (10 mL). The ammonia solution was concentrated under vacuum and the residue purified by preparative HPLC to afford the desired compound (67 mg, 40 %); ¹H NMR (300 MHz, ^methanol) δ 8.51 (br s, 1 H), 8.12 (d, J = 8.8 Hz, 2H), 7.77 (d, J = 8.8 Hz, 2H), 7.26 - 7.16 (m, 4H). 4.1 1 (m, 2H), 3.57 - 3.47 (m, 2H), 3.44-3.35 (m, 2H), 3.25 - 3.1 1 (m, 3H), 3.10 -2.99 (m, 1 H), 2.69 (s, 3H). LCMS Method-D: RT 5.42 min; m/z 476.19 [M+H]\

Example 4

General Procedure A

I4

A solution of appropriate carboxylic acid (0.220 mmol - 0.590 mmol, 1 equiv.) in DCM (2-10 mL) containing Λ/,/V-diisopropylethylamine (3 equiv.), N-ethyl-N'-(3- dimethylaminopropyl)carbodiimide hydrochloride (1.3 equiv.) and 1 -hydroxybenzotriazole hydrate (1.3 equiv.) was stirred for 20 minutes. (S -1-amino-3-((2,3-dihydro-1 H-inden-2- yl)amino)propan-2-ol I4 (0.9 equiv.) in DCM (1 mL) was added and the mixture was stirred at room temperature for 24 hours under a nitrogen atmosphere. Water (15 mL) and DCM (15 mL) were added. The aqueous phase was extracted with DCM (2 15 mL) and the combined organic extracts were washed with brine (20 mL), dried (sodium sulfate), filtered and concentrated in vacuo. The residue was dissolved in methanol and applied to an SCX cartridge (1 g). The cartridge was washed with methanol (10 mL) and eluted with 2 M ammonia in ethanol (10 mL). The ammonia solution was concentrated under vacuum and the residue purified by preparative-HPLC (Waters) to afford the desired compound.

General Procedure B

I6 A solution of appropriate carboxylic acid (0.209 - 0.889 mmol, 1 equiv.) in DCM (2-10 mL) containing A/./V-diisopropylethylamine (3 equiv.), A/-ethyl-A/'-(3- dimethylaminopropyl)carbodiimide hydrochloride (1.3 equiv.) and 1 -hydroxybenzotriazole hydrate (1.3 equiv.) was stirred for 20 minutes. (S) -1 -Amino-3-((2.3-dihydro-1 H-inden-2- yl)amino)propan-2-ol I6 (0.9 equiv.) in DCM (1 mL) was added and the mixture was stirred at room temperature for 24 hours under a nitrogen atmosphere. Water (10 mL) and DCM (10 mL) were added. The aqueous phase was extracted with DCM (2 10 mL) and the combined organic extracts were washed with brine (15 mL), dried (sodium sulfate), filtered and concentrated in vacuo. The residue was dissolved in methanol and applied to an SCX cartridge (1 g). The cartridge was washed with methanol (10 mL) and eluted with 2 M ammonia in ethanol (10 mL). The ammonia solution was concentrated under vacuum and the residue purified by preparative-HPLC (Waters) to afford the desired compound.

18 LCMS B

Method-D:

o

RT 4.57 Reaction

(S)-/V-(3-((2,3-dihydro-1 H-inden-2- min; m/z solvent:

77.0mg, 0.353

yl)amino)-2-hydroxypropyl)-4-((1- 407.4 DCM (7.0 mmol

methyl-1 H-pyrazol-4- [M+H]⁺. mL) yl)oxy)benzamide

Example 5: 4-{(3-((2,3-Dihydro-1 H-inden-2-yl)amino)-2- hydroxypropyl)carbamoyl)benzoic acid (19)

19

(a) Methyl 4-((3-((2, 3-di hydro- 1 H-inden-2-yl)amino)-2-hydroxypropyl)carbamoyl)benzoate

(A3)

A solution of 4-(methoxycarbonyl)benzoic acid (1 .95 g, 10.8 mmol) in DCM (15 mL) containing Λ/,/V-diisopropylethylamine (1 1.5 mL, 66.2 mmol), N-ethyl-N'-(3- dimethylaminopropyl)carbodiimide hydrochloride (5.50 g, 28.7 mmol) and 1- hydroxybenzotriazole hydrate (4.39 g, 28.7 mmol) was stirred for 20 minutes. 1-Amino-3- ((2.3-dihydro-1 H-inden-2-yl)amino)propan-2-ol (I2) (2.10 g, 10.2 mmol) in DCM (1 mL) was added and the mixture was stirred at room temperature for 24 hours under a nitrogen atmosphere. Water (10 mL) and DCM (10 mL) were added. The aqueous phase was extracted with DCM (2 10 mL) and the combined organic extracts were washed with brine (15 mL), dried (sodium sulfate), filtered and concentrated in vacuo. The residue was dissolved in methanol (7 mL) and loaded on to an SCX cartridge. The cartridge was washed with methanol (30 mL) and eluted with 2 M ammonia in ethanol (20 mL). The ammonia solution was concentrated under vacuum and the residue was purified by column chromatography (S1O2, 0-30% ethyl methanol/DCM) to give desired compound (644 mg, 16 %): LCMS Method-D: RT 4.57 min; m/z 369.3 [M+H]⁺.

(b) 4-((3-((2,3-Dihydro- 1 H -inden-2-yl)amino)-2-hydroxypropyl)carbamoyl)benzoic acid (19) To a solution of methyl 4-((3-((2.3-dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)carbamoyl) benzoate A3 (481 mg, 1 .31 mmol) in THF (15 mL) and water (6 mL) was added LiOH (47 mg, 1.96 mmol) in water (1 mL). The reaction was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure and dissolved in water (2 mL). The material was applied to a desalting column (1 Og, Phenomenex, Strata-XL

Polymeric reversed phase). The column was washed with water (20 mL) and eluted with methanol (20 mL). The methanol eluate was concentrated to give the desired compound (395 mg, 85%). LCMS Method-D: RT 4.17 min; m/z 355.4 [M+H]\

Example 6: 4-(Azetidine-1 -carbonyl)-A -(3-((2,3-dihydro-1 H-inden-2-yl)ami

hydroxypropyl)benzamide (20)

19

A solution of 4-((3-((2,3-dihydro-1 /-/-inden-2-yl)amino)-2-hydroxypropyl)carbamoyl)benzoic acid 19 (95 mg, 0.27 mmol) in DCM (7 mL) containing A/,/V-diisopropylethylamine (201 μί, 1.15 mmol), A/-ethyl-A/'-(3-dimethylaminopropyl)carbodiimide hydrochloride (67 mg, 0.35 mmol) and 1 -hydroxybenzotriazole hydrate (54 mg, 0.35 mmol) was stirred for 20 minutes. Azetidine (18 L, 0.27 mmol) was added and the mixture was stirred at room temperature for 24 hours under a nitrogen atmosphere. Water (10 mL) and DCM (10 mL) were added. The aqueous phase was extracted with DCM (2 10 mL) and the combined organic extracts were washed with brine (15 mL), dried (sodium sulfate), filtered and concentrated in vacuo. The residue was purified by preparative HPLC to give the desired compound (32 mg, 30 %); ¹H NMR (300 MHz, ^-methanol) δ 8.43 (br s, 1 H), 7.95 (m, 2H), 7.73 (m, 2H), 7.29 - 7.19 (m, 4H), 4.38 (t, J = 7.7 Hz, 2H), 4.24 - 4.08 (m, 4H), 3.61 - 3.50 (m, 2H), 3.47 - 3.38 (m, 2H), 3.29 - 3.13 (m, partially obscured by solvent), 3.10 - 3.03 (m, 1 H). LCMS Method-D: RT 4.22 min; m/z 394.4 [M+H]\ Example 7: A -(3-((2,3-Dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)-4-(pyrrolidine-1 - carbonyl)benzamide (21 )

19

A solution of 4-((3-((2,3-dihydro-1 /- -inden-2-yl)amino)-2-hydroxypropyl)carbamoyl)benzoic acid 19 (102 mg, 0.29 mmol) in DCM (7 mL) containing Λ/,/V-diisopropylethylamine (201 L, 1.15 mmol), N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide hydrochloride (72 mg, 0.37 mmol) andl -hydroxybenzotriazoie hydrate (57 mg, 0.37 mmol) was stirred for 20 minutes. Pyrrolidine (24 μΙ_, 0.29 mmol) was added and the mixture was stirred at room temperature for 24 hours under a nitrogen atmosphere. Water (10 mL) and DCM (10 mL) were added. The aqueous phase was extracted with DCM (2 10 mL) and the combined organic extracts were washed with brine (15 mL), dried (sodium sulfate), filtered and concentrated in vacuo. The material was dissolved in DCM (7 mL) containing A/,A/-diisopropylethylamine (201 L, 1.15 mmol), N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide hydrochloride (72 mg, 0.37 mmol) and 1 -hydroxybenzotriazoie hydrate (57 mg, 0.37 mmol). The mixture was stirred for 20 minutes. Pyrrolidine (1 19 μί, 1 .44 mmol) was added and the mixture was stirred at room temperature for 24 hours under a nitrogen atmosphere. Water (10 mL) and DCM (10 mL) were added. The aqueous phase was extracted with DCM (2 x 10 mL) and the combined organic extracts were washed with brine (15 mL), dried (sodium sulfate), filtered and concentrated in vacuo. The residue was purified by preparative HPLC to give the desired compound (7 mg, 6 %); Ή NMR (300 MHz, ^-methanol) δ 8.54 (br s, 1 H), 7.94 (d, J = 8.1 Hz, 2H), 7.63 (d, J = 8.0 Hz, 2H), 7.29 - 7.19 (m, 4H), 4.1 1 (m, 2H), 3.64 (m, 2H), 3.53 (m, 2H), 3.47 - 3.38 (m, 4H), 3.28 - 3.00 (m, 4H), 2.09 - 1 .89 (m, 4H). LCMS Method-D: RT 4.37 min; m/z 408.4 [M+Hf. Example 8: A -(3-((2,3-Dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)-4-(piperidine-1 - carbon l)benzamide (22)

19 A solution of 4-((3-((2,3-dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)carbamoyl)benzoic acid 19 (105 mg, 0.296 mmol) in DCM (7 mL) containing A/./V-diisopropylethylamine (206 μΙ_, 1.19 mmol), A/-ethyl-A/'-(3-dimethylaminopropyl)carbodiimide hydrochloride (74 mg, 0.39 mmol) and 1 -hydroxybenzotriazole hydrate (59 mg, 0.39 mmol) was stirred for 20 minutes. Piperidine (29 μΙ_, 0.30 mmol) was added and the mixture was stirred at room temperature for 24 hours under a nitrogen atmosphere.Water (10 mL) and DCM (10 mL) were added. The aqueous phase was extracted with DCM (2 10 mL) and the combined organic extracts were washed with brine (15 mL), dried (sodium sulfate), filtered and concentrated in vacuo. The residue was purified by preparative HPLC to give the desired compound (76 mg, 61 %). ¹H NMR (300 MHz, ^-methanol) 5 8.39 (br s, 1 H), 7.95 (m, 2H), 7.51 (m, 2H), 7.29 - 7.19 (m, 4H), 4.14 (m, 2H), 3.74 (m, 2H), 3.62 - 3.51 (m, 2H), 3.49 - 3.42 (m, 2H), 3.40 - 3.37 - (m, 2H), 3.29 - 3.13 (m, 3H), 3.10 - 3.03 (m, 1 H), 1 .72 (br m, 4H), 1.56 (br m, 2H); LCMS Method-D: RT 4.58 min; m/z 422.3 [M+H]'. Example 9: Af-(3-((2,3-Dihydro-1 H-inden-2-yI)amino)-2-hydroxypropyI)-[1 ,1'-biphenyI]- 4-carboxamide (24)

A solution of 4-biphenylcarboxylic acid (0.074 g, 0.37 mmol) in DCM (4 mL) containing N,N- diisopropylethylamine (0.132 mL, 1.01 mmol), /V-ethyl-/V-(3- dimethylaminopropyl)carbodiimide hydrochloride (0.098 g, 0.51 mmol) and 1 - hydroxybenzotriazole hydrate (wetted with not less than 20 wt. % water,0.098 g, 0.509 mmol) was stirred at room temperature for 20 minutes. A solution of 1-amino-3-((2,3-dihydro- 1 H-inden-2-yl)amino)propan-2-ol I2 (0.070 g, 0.339 mmol) in DCM (2 mL) was added. The mixture was then stirred at room temperature for 20 hours whilst under a nitrogen

atmosphere. Water was added and the resulting mixture formed a white precipitate which was filtered and dried. The white solid was washed with diethyl ether and collected by filtration to give the desired compound as an amorphous white solid (0.105 g, 80%). LCMS Method-D: RT 5.28 min; m/z 387.4 [M+H]\ Example 10: A -{3-((2,3-Dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)-[1 ,1'-biphenyl]- 3-carboxamide 25)

A solution of 3-biphenylcarboxylic acid (0.074 g, 0.37 mmol) in DCM (4 mL) containing N,N- diisopropylethylamine (0.132 mL, 1.01 mmol). A/-ethyl-/V-(3- dimethylaminopropyl)carbodiimide hydrochloride (0.098 g, 0.51 mmol) and 1 - hydroxybenzotriazole hydrate (wetted with not less than 20 wt. % water, 0.098 g, 0.51 mmol) was stirred at room temperature for 20 minutes. A solution of 1 -amino-3-((2.3-dihydro-1 H- inden-2-yl)amino)propan-2-ol I2 (0.070 g, 0.34 mmol) in DCM (2 mL) was added. The mixture was stirred at room temperature for 20 hours under a nitrogen atmosphere. Water and DCM were added. The aqueous phase was extracted with DCM (2 15 mL) and the combined organic extracts were washed with brine, dried (magnesium sulfate), filtered and concentrated under reduced pressure. The product was purified by column chromatography (12 g SiO? cartridge, 0-10% methanol in DCM) to give the desired product as an amorphous white solid (0.026 g, 19%). LCMS Method-D: RT 5.28 min; m/z 387.4 [M+H]' .

Example 11 : AI-(3-((2,3-Dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)-[1 ,1^!-biphenyl]- 2-carboxamide (26)

A solution of 2-biphenylcarboxylic acid (0.074 g, 0.373 mmol) in DCM (4 mL) containing N,N- diisopropylethylamine (0.132 mL, 1.01 mmol, 3 equiv.), A/-ethyl-/V-(3- dimethylaminopropyl)carbodiimide hydrochloride (0.098 g, 0.509 mmol, 1 .5 equiv) and 1 - hydroxybenzotriazole hydrate (wetted with not less than 20 wt. % water,0.098 g, 0.509 mmol) was stirred at room temperature for 20 minutes. After this time, a solution of 1-amino- 3-((2,3-dihydro-1 H-inden-2-yl)amino)propan-2-ol I2 (0.070 g, 0.339 mmol) in DCM (2 mL) was added. The mixture was then stirred at room temperature for 20 hours whilst under a nitrogen atmosphere. Water and DCM were added. The aqueous phase was extracted with DCM (2 15 mL) and the combined organic extracts were washed with brine, dried

(magnesium sulfate), filtered and concentrated under reduced pressure. The product was purified by column chromatography (12 g SiO? Cartridge, 0-10% methanol in DCM) to give the desired product as an amorphous solid (0.060, 45%). LCMS Method-D: RT 4.92 min; m/z 387.3 [M+H] \ Example 12: Ai-(3-((2,3-DihycSro-1 H-inden-2-yI)amino)-2-hydroxypropyI)-2-fIuoro-4- (mor holine-4-carbonyl)benzamide (27)

(a) (4-Bromo-3-fluorophenyl)(morpholino)methanone (A6)

4-Bromo-3-fluorobenzoic acid (2.19 g, 10.0 mmol), morpholine (2.59 mL, 30.0 mmol), DMAP (122 mg, 10 mol%) and EDCI.HCI (2.88 g, 15.0 mmol) in DCM (100 mL), were stirred at room temperature. After 17 hours, the mixture was added to 2% w/v aqueous sodium hydroxide (100 mL) and the separated aqueous phase was extracted with DCM (2 50 mL). The combined organic extracts were washed with 1 M HCI (75 mL), brine (50 mL), dried over sodium sulfate and concentrated. Chromatography (40 g silica cartridge, 0-60% ethyl acetate/petroleum benzine 40-60 °C) gave the desired compound as a colourless syrup

(2.63 g, 91 %): Ή NMR (400 MHz, CDCb) δ 7.61 (dd, J = 8.1 , 6.8 Hz, 1 H), 7.19 (dd, J = 8.5, 1.9 Hz, 1 H), 7.08 (ddd, J = 8.1 , 1 .9, 0.7 Hz, 1 H), 3.70 (br s, 6H), 3.45 (br s, 2H); LCMS-B: RT 3.49 min; m/z 290.0, 288.1 [M+H] \ (b) Methyl 2-fluoro-4-(morpholine-4-carbonyl)benzoate (A7)

(4-Bromo-3-fluorophenyl)(morpholino)methanone A6 (2.62 g, 9.09 mmol), dry methanol (20 mL), PdCb(dppf).DCM (376 mg, 5 mol%) and triethylamine (2.54 mL, 18.2 mmol) were loaded into a Schlenk tube and flushed with nitrogen. The tube was flushed with carbon monoxide and the mixture brought to reflux under carbon monoxide (balloon). After 18 hours, the mixture was cooled to room temperature, filtered through Celite and the Celite washed with methanol (40 mL). The combined filtrates were concentrated and

chromatography (40 g silica cartridge, 20-60% ethyl acetate/petroleum benzine 40-60 C) gave the desired compound as a pale orange solid (2.25 g, 93%).¹H NMR (400 MHz, CDCb) δ 7.99 (t, J = 7.3 Hz, 1 H), 7.25 - 7.17 (m, 2H), 3.94 (s, 3H), 3.78 (br s, 4H), 3.63 (br s, 2H), 3.40 (br s, 2H); LCMS-A: RT 5.30 min; m/z 268.1 [M+H]\

(c) 2-Fluoro-4-(morpholine-4-carbonyl)benzoic acid (Ad)

Methyl 2-fluoro-4-(morpholine-4-carbonyl)benzoate A7 (1.00 g, 3.74 mmol) was dissolved in THF (20 mL) and a solution of lithium hydroxide monohydrate (188 mg, 4.49 mmol) in water (10 mL) was added. The mixture was stirred vigorously at room temperature. After two hours, the volatile solvents were removed in vacuo. The aqueous residue was cooled to 4 C and 3.0 aqueous HCI (5 mL) cooled to 4 °C was added. The resulting slurry was diluted with water (5 mL) and filtered. The collected solids were washed with water (5 mL) and air dried to give the desired compound as a white solid (817 mg, 86%).¹H NMR (400 MHz, 4- methanol ) δ 8.05 - 7.99 (m, 1 H), 7.32 (s, 1 H), 7.30 (dd, J = 3.7, 1 .4 Hz, 1 H), 3.76 (br s, 4H), 3.63 (br s, 2H), 3.42 (br s, 2H); LCMS-B: RT 3.20 min; m/z 254.2 [M+H]'; 276.2 [M+Na]\ (d) N -(3-((2,3-Dihydro-1H-inden-2-yl)amino)-2-hydroxypropyl)-2-fluoro-4-(morpholine-4- carbonyl)benzamide (27)

A mixture of 2-fluoro-4-(morpholine-4-carbonyl)benzoic acid A8 (0.057 g, 0.23 mmol) and triethylamine (0.14 mL, 0.97 mmol) in DCM (5 mL) was cooled to 0 °C under a nitrogen atmosphere and isobutyl chloroformate (32 L, 0.24 mmol) was added dropwise. The mixture was stirred for 45 minutes at 0 °C and allowed to settle. The liquid phase was transferred via syringe to a solution of 1 -amino-3-((2,3-dihydro-1 /-/-inden-2-yl)amino)propan- 2-ol I2 (0.050 g, 0.24 mmol) in DCM (10 mL) at 0 °C. After 1 .25 hours, the mixture was diluted with DCM (10 mL) and aqueous sodium hydroxide (2 M, 25 mL) and the separated aqueous phase was extracted with DCM (2 25 mL). The combined organic extracts were washed with brine (25 mL), dried (MgSC ) and concentrated in vacuo. The residue was purified by column chromatography (24 g SiO? cartridge, 0-10% MeOH (containing 1 % v/v 2 M NH3 in MeOH) in DCM) to give a white solid. The solid was dissolved in a minimal amount of DCM and the product was precipitated by the addition of cyclohexane. The precipitate was vacuum filtered and air dried to give the desired compound as a white solid (0.020 g, 20%). H NMR (400 MHz, cf₆-DMSO) δ 8.46 (t, J = 5.7 Hz, 1 H), 7.69 (t, J = 7.5 Hz, 1 H), 7.42 - 7.35 (m, 1 H), 7.35 - 7.29 (m, 1 H), 7.25 - 7.19 (m, 2H), 7.18 - 7.12 (m, 2H), 5.38 (s, 1 H), 3.95 - 3.74 (m, 2H), 3.71 - 3.48 (m, 6H), 3.23 - 3.10 (m, 2H), 3.00 - 2.82 (m, 3H), 2.81 - 2.69 (m, 1 H), residual H2O obscured peaks in the a Iky I region; LCMS-B: RT 3.20 min; m/z 442.3 [M+H] '. Example 13: A -(3-((5-Bromo-2,3-dihydro-1 «-inden-2-yS)amino)-2-hydroxypropyS)-4- chlorobenzamide 28)

A10 28

(a) teti- Butyl (3-(4-chlorobenzamido)-2-hydroxypropyl)carbamate (A9)

A solution of 4-chlorobenzoic acid (3.00 g, 19.2 mmol) in DCM (40 mL) containing N,N- diisopropylethylamine (10.01 mL, 57.5 mmol, ), V-ethyl-/V-(3- dimethylaminopropyl)carbodiimide hydrochloride (4.77 g, 25.0 mmol) and 1- hydroxybenzotriazole hydrate (wetted with not less than 20 wt. % water,4.76 g, 25.0 mmol) was stirred at room temperature for 20 minutes. /V-Boc-1 ,3-diamino-2-propanol (4.01 g, 21 .1 mmol) was added and the mixture was stirred at room temperature for 20 hours under a nitrogen atmosphere. Water was added and the aqueous phase was extracted with DCM (2 20 mL). The combined organic extracts were washed with brine, dried (sodium sulfate), filtered and concentrated under reduced pressure. The material was purified using column chromatography (120 g S1O2 Cartridge, 0-30% methanol in DCM) to give of the desired compound as a white foam (3.44 g, 54%). ¹H NMR (300 MHz, cfe-DMSO) δ 8.47 (t, J=5.50 Hz, 1 H), 7.88 (d, J=8.58 Hz, 2H), 7.54 (d, J=8.58 Hz, 2H), 6.70 (t, J=5.58 Hz, 1 H), 4.93 (d, J=5.06 Hz, 1 H), 3.66-3.61 (m, 1 H), 3.30 - 3.26 (m, 1 H), 3.26 - 3.1 1 (m. 1 H), 3.04 - 2.50 (m, 2H), 1 .38 (s, 9H). (b) N -(3 -A mi no -2-hydroxypropyl) -4 -chlorobenzamide hydrochloride (A 10)

A solution of iert-butyl (3-(4-chlorobenzamido)-2-hydroxypropyl)carbamate A9 (3.44 g, 10.5 mmol) in 4.0 M hydrochloric acid in 1.4-dioxane (10.4 mL, 41 .9 mmol) and 1 ,4-dioxane (15 mL) was heated at 50 °C for 2.5 hours. The solvent was evaporated under reduced pressure to give the desired compound product as a white foam (2.77 g, 99%). ¹ H NMR (300 MHz, afe- DMSO) δ 8.80 (t, J=5.61 Hz, 1 H), 7.91 (d, J=6.78 Hz, 2H), 7.55 (d, J=8.55 Hz, 2H), 3.95 - 3.82 (m, 1 H), 3.57 - 3.23 (m, 5H), 3.02 - 2.85 (m, 1 H), 2.77 - 2.60 (m, 1 H). (c) N -(3-((5-Bromo-2, 3-dihydro- 1 H -inden-2-yl)amino)-2-hydroxypropyl)-4-chlorobenzamide

(28)

To a solution of A/-(3-amino-2-hydroxypropyl)-4-chlorobenzamide hydrochloride A10 (0.070 g, 0.26 mmol) in 1 ,2-dichloroethane (1 mL) and methanol (1 mL) was added N,N- diisopropylethylamine (0.046 mL, 0.26 mmol), 5-bromo-1 H-inden-2(3H)-one (0.067 g, 0.32 mmol), acetic acid (0.024 mL, 0.40 mmol), and sodium triacetoxyborohydride (0.224 g, 1.05 mmol). The reaction mixture was stirred at room temperature for 20 hours under a nitrogen atmosphere. The reaction mixture was diluted with 2.5 M aqueous sodium hydroxide and DCM. The aqueous phase was extracted with DCM (2 15 mL) and the combined organic extracts were washed with brine, dried (sodium sulfate), filtered and concentrated under reduced pressure. The material was purified by silica column chromatography (12 g S\0? Cartridge, 0-15% methanol in DCM) to give the desired compound as a light brown solid (0.040 g, 35%): LCMS RT 4.41 min; m/z 424.7 [M+H]\

Example 14: 4-C loro-/V-(4-((2,3-dihydro-1 H-inden-2-yl)ami

h roxybutyl)benzamide (29)

(a) 2-(But-3-en- 1 -yl)isoindoline-1 ,3-dione (A 1 1)

A mixture of potassium phthalimide (500 mg, 2.70 mmol), acetonitrile (5 mL) and 4- bromobut-1-ene (0.548 mL, 5.40 mmol) was stirred at 90 °C. After 18 hours, the mixture was cooled to room temperature, filtered and the collected solids were washed with ethyl acetate

(5 mL). The combined filtrates were concentrated and column chromatography (12 g silica cartridge, 0-100% ethyl acetate/hexanes) gave the desired compound as a white solid (431 mg, 79%).¹H NMR (400 MHz, CDCI₃) δ 7.87 - 7.80 (m, 2H), 7.73 - 7.67 (m, 2H), 5.86 - 5.72 (m, 1 H), 5.1 1 - 4.99 (m, 2H), 3.77 (t, J = 7.1 Hz, 2H), 2.49 - 2.40 (m, 2H).LCMS-B: RT 3.71 min; m/z 202.1 [M+H] '.

(b) 2-(2-(Oxiran-2-yl)ethyl)isoindoline- 1,3-dione (A 12)

A mixture of 2-(but-3-en-1-yl)isoindoline-1 ,3-dione A1 1 (428 mg, 2.13 mmol), chloroform (5 mL) and 70-75% mCPBA (629 mg, 2.55 mmol) was stirred at room temperature. After 18 hours, the mixture was quenched with 10% w/v aqueous sodium thiosulfate (2.5 mL) and was stirred vigorously for 5 minutes. The mixture was diluted with a saturated aqueous solution of sodium bicarbonate (5 mL), water (20 mL) and chloroform (10 mL). The separated aqueous phase was extracted with chloroform (2 x 10 mL). The combined organic extracts were dried over sodium sulfate and concentrated in vacuo to give the desired compound as a white solid (451 mg, 98%). The material was carried forward without further purification. Ή N R (400 MHz, CDC ) δ 7.88 - 7.82 (m, 2H), 7.74 - 7.69 (m, 2H), 3.97 - 3.80 (m, 2H), 3.03 - 2.94 (m, 1 H), 2.74 - 2.68 (m, 1 H), 2.47 - 2.41 (m, 1 H), 2.04 - 1.93 (m, 1 H), 1 .90 - 1 .79 (m, 1 H).LCMS-B: RT 3.43 min; m/z 218.1 [M+H]^*

(c) 2-(4-((2.3-Dihydro- 1 H-inden-2-yl)amino)-3-hydroxybutyl)isoindoline- 1,3-dione (A 13) A mixture of 2-(2-(oxiran-2-yl)ethyl)isoindoline-1 ,3-dione A12 (100 mg, 0.460 mmol), dry acetonitrile (2 mL), 2-aminoindane (64 mg, 0.48 mmol) and calcium triflate (78 mg, 50 mol%) was stirred at room temperature. After two hours, the mixture was concentrated in vacuo and purified by column chromatography (12g S\0? cartridge, 0-20% methanol/DCM) to give the desired compound as a colourless solid (139 mg, 86%). ¹H NMR (400 MHz. DMSO) δ 7.92 - 7.77 (m, 2H), 7.32 - 7.1 1 (m, 6H), 4.04 - 3.96 (m, 1 H), 3.28 (dd, J = 16.6, 7.7 Hz, overlaps with solvent), 2.92 (dd, J = 16.6, 5.1 Hz, 2H); LCMS-B: m/z 351 .2 [M+H]^*.

(d) 4-Amino- 1 -((2,3-dihydro-1H-inden-2-yl)amino)butan-2-ol (A 14)

A mixture of 2-(4-((2,3-dihydro-1 H-inden-2-yl)amino)-3-hydroxybutyl)isoindoline-1 ,3-dione A13 (137 mg, 0.391 mmol), ethanol (2 mL) and hydrazine hydrate (0.2 mL) was stirred at 80 C. After three hours, the mixture was cooled to room temperature, the resulting slurry filtered and the collected solids washed with cold ethanol (2 mL). The combined filtrates were concentrated to give the title compound as a white solid (101 mg, >100 % yield). The material was carried forward without further purification.

(e) 4-Chloro-N-(4-((2,3-dihydro- 1H-inden-2-yl)amino)-3-hydroxybutyl)benzamide (29) A mixture of 4-amino-1-((2,3-dihydro-1 /-/-inden-2-yl)amino)butan-2-ol A14 (44 mg, 0.20 mmol), MeCN (2 mL), DIPEA (0.070 mL, 0.40 mmol) and 4-chlorobenzoyl chloride (0.026 mL, 0.20 mmol) was stirred at room temperature. After 18 hours, methanol (0.1 mL) was added and the mixture loaded onto a 5 g SCX cartridge. The cartridge was washed with methanol (40 mL) then eluted with 3.5 M ammonia in methanol (40 mL). The basic eluate was concentrated in vacuo, and preparative TLC (5% methanol/DCM) gave the desired compound as a colourless syrup (7 mg, 9% yield). ¹H NMR (400 MHz, ^-methanol) δ 7.83 - 7.78 (m, 2H), 7.49 - 7.45 (m, 2H). 7.21 - 7.10 (m, 4H), 3.86 - 3.76 (m, 1 H), 3.75 - 3.64 (m, 1 H), 3.61 - 3.44 (m, 2H), 3.26 - 3.16 (m, 2H), 2.88 - 2.79 (m, 3H), 2.76 - 2.67 (m, 1 H), 1.87 - 1 .77 (m, 1 H), 1 .75 - 1 .64 (m, 1 H). Example 15: 4-Chloro-/V-{2-hydroxy-3-{(1 ,2,3,4-tetrahydronaphthalen-2- yl)amino)propy!)benzamide (30)

A16 30

(a) 2-(2-Hydroxy-3-((1,2,3,4-tetrahydronaphthalen-2-yl)amino)propyl)isoindoline- 1,3-dione

(A 15)

A mixture of A/-(2,3-epoxypropyl)phthalimide (0.471 g, 2.31 mmol) and 1 ,2,3.4-tetrahydro-2- naphthalenamine (0.310 g, 2.10 mmol) in ethanol (3 mL) was heated at reflux for 20 hours under a nitrogen atmosphere. The volatiles were removed in vacuo. The material was adsorbed directly onto silica gel and the product was purified by column chromatography (12 g S1O2 Cartridge, 0-20% methanol in DCM) to give the desired compound as a light yellow oil.

(0.267 g, 36%). LCMS-D: RT 4.95 min; m/z 351.2 [M+H] \

(b) 1-Amino-3-((1,2,3,4-tetrahydronaphthalen-2-yl)amino)propan-2-ol (A 16)

A mixture of 2-(2-hydroxy-3-((1 ,2,3,4-tetrahydronaphthalen-2-yl)amino)propyl)isoindoline- 1.3-dione A15 (0.267 g, 0.762 mmol) and hydrazine monohydrate (0.075 mL, 1 .5 mmol) in ethanol (2 mL) and DCM (1 mL) was stirred at room temperature for 20 hours under a nitrogen atmosphere. The volatiles were removed by rotary evaporation. The residue was dissolved in methanol and passed through a SCX cartridge (5g, Agilent, Bond Elut, Mega BE-SCX) washing with 100% methanol and eluting with 100% 2 M ammonia in methanol solution to give the desired compound (0.150 g, 89%). The material was used in the next step without further purification. (c) 4-Chloro- -(2-hydroxy-3-((1 ,2,3A-tetrahydronaphthalen-2-yl)amino)propyl)benzamide

(30)

A solution of 4-chlorobenzoic acid (0.1 17 g, 0.749 mmol) in /V,A/-dimethylformamide (2 mL) containing Λ/,/V-diisopropylethylamine (0.474 mL, 2.72 mmol), /V-ethyl-/V-(3- dimethylaminopropyl)carbodiimide hydrochloride (0.196 g, 1.02 mmol) and 1 - hydroxybenzotriazole hydrate (wetted with not less than 20 wt. % water, 0.156 g, 1.02 mmol) was stirred at room temperature for 20 minutes under a nitrogen atmosphere. A solution of 1-amino-3-((1 ,2,3.4-tetrahydronaphthalen-2-yl)amino)propan-2-ol A16 (0.150 g, 0.681 mmol) in A/,A/-dimethylformamide (2 mL) was added and stirred at room temperature for 20 hours under a nitrogen atmosphere. Water and ethyl acetate were added. The aqueous phase was extracted with ethyl acetate (2 20 mL) and the combined organic extracts were washed with water (*3), brine, dried (sodium sulfate), filtered and concentrated in vacuo to give the product. The material was adsorbed directly onto silica gel and purified by column chromatography (12 g S1O2 Cartridge, 0-20% DCM in methanol) to give the desired compound as an amorphous white solid (0.075 g, 30%) ¹ H MR (300 MHz, (^-methanol) δ 7.83 (d, J=8.55 Hz, 2H), 7.49 (d, J=8.55 Hz. 2H), 7.06 (br. s., 4H), 3.99 - 3.95 (m, 1 H), 3.50 - 3.48 (m, 2H), 3.10 - 2.58 (m, 7H), 2.15 - 2.1 1 (m, 1 H), 1.68 - 1.61 (m, 1 H). LCMS-D: RT 3.63 min; m/z 359.3 [M+H]\

Example 16: 4-Chloro-/V-{3-{(7-fluoro-1 ,2,3,4-tetrahydronaphthalen-2-yl)amino)-2- h droxypropyl)benzamide (31 )

A18 31 (a) \ert- Butyl (3-((7-fluoro- 1.2,3,4-tetrahydronaphthalen-2-yl)amino)-2- hydroxypropyl)carbamate (A 17)

7-Fluoro-1 ,2.3,4-tetrahydro-napthalene-2-ylamine hydrochloride (120 mg) was neutralised by adding 10% w/v NaOH (aq.) (5 mL) and DCM (5 mL) to the salt. The phases were separated and the aqueous phase was extracted with DCM (3 x 10 mL). The combined organic phases were dried with sodium sulfate and concentrated under reduced pressure to give the free amine (76 mg) as an oil. To a solution of 7-fluoro-1 ,2,3,4-tetrahydro-napthalene-2-ylamine (76 mg, 0.46 mmol) in dry acetonitrile (1 .5 mL) was added tert-butyl A/-(oxiran-2-yl methyl) carbamate (159 mg, 0.92 mmol). The flask was sealed and the reaction was allowed to stir at 80 C overnight. The reaction mixture was concentrated under reduced pressure and the residue was dissolved in methanol and applied to an SCX cartridge (5 g). The cartridge was washed with methanol (25 mL) and eluted with 2 M ammonia in methanol (15 mL). The volatiles were removed under vacuum to afford the desired compound (131 mg, 84%). The material was taken onto the next step without further purification.

(b) 1 -Amino-3-((7-fluoro- 1,2,3, 4-tetrahydronaphthalen-2-yl)amino)propan-2-ol (A 18) ie/ -Butyl (3-((7-fluoro-1 ,2,3.4-tetrahydronaphthalen-2-yl)amino)-2-hydroxypropyl)carbamate A17 (131 mg, 0.39 mmol) was dissolved in DCM (7 mL) and to the solution was added TFA (0.531 mL, 7.74 mmol). The reaction was stirred at room temperature under a nitrogen atmosphere overnight and the solvent was removed under reduced pressure. The resulting oil was dissolved in methanol (7 mL) and applied to an SCX cartridge. The cartridge was washed with methanol (25 mL) and eluted with 2 M ammonia in ethanol (15 mL). The ammonia eluate was concentrated under vacuum to give the desired compound (91 mg, 99 %); ¹ H NMR (300 MHz, CDCI₃) δ 7.01 (m, 1 H), 6.82 - 6.72 (m, 2H), 3.68 (m, 1 H), 3.02 - 2.50 (m, 1 1 H), 2.01 (m, 1 H), 1 .59 (m, 1 H).

(c) 4-Chloro-N -(3-((7-fluoro- 1 ,2,3,4-tetrahydronaphthalen-2-yl)amino)-2- hydroxypropyl)benzamide (31)

To a solution of 4-chlorobenzoic acid (66 mg, 0.42 mmol) in DCM (7.0 mL) was added N,N- diisopropylethylamine (221 L, 1.27 mmol), A/-ethyl-/V-(3-dimethylaminopropyl)carbodiimide hydrochloride (106 mg, 0.552 mmol) and 1 -hydroxybenzotriazole hydrate (84.5 mg, 0.552 mmol) under a nitrogen atmosphere. The solution was stirred for 20 minutes at room temperature and 1-amino-3-((7-fluoro-1 ,2,3,4-tetrahydronaphthalen-2-yl)amino)propan-2-ol A18 (91 mg, 0.38 mmol) was added. The reaction was stirred at room temperature for 24 hours. Water (25 mL) and DCM (25 mL) were added and the aqueous phase was extracted with DCM (3 20 mL). The combined organic extracts were washed with water (50 mL), brine (50 mL), dried over sodium sulfate and concentrated. The resulting oil was dissolved in methanol and applied to an SCX cartridge (5 g), the cartridge was washed with methanol (30 mL) then eluted with 2 M ammonia in ethanol (20 mL). The ammonia eluate was

concentrated and the resulting residue was purified by preparative HPLC (Waters) to give the desired compound (5 mg, 3 %). LCMS-D RT 5.17 min; m/z 377.3.

Example 17: 4-Chloro-/V-{3-{(6-fluoro-1 ,2,3,4-tetrahydronaphthalen-2-yl)amino)-2- hydr xypropyl)benzamide (32)

(a) tert-Butyl (3-((6-fluoro- 1,2,3.4-tetrahydronaphthalen-2-yl)amino)-2- hydroxypropyl)carbamate (A 19)

6-Fluoro-1 ,2,3.4-tetrahydro-napthalene-2-ylamine hydrochloride (100 mg) was neutralised by adding 10% NaOH (aq.) (5 mL) and DCM (10 mL) to the salt. The phases were separated and the aqueous phase was extracted with DCM (2 10 mL). The combined organic phases were dried with sodium sulfate and concentrated under reduced pressure to give the free amine as an oil (86 mg). To a solution of 6-fluoro-1 ,2,3,4-tetrahydro-napthalene-2-ylamine (86 mg, 0.52 mmol) in dry acetonitrile (1 .5 mL) was added ferf-butyl A/-(oxiran-2-yl methyl) carbamate (180 mg, 1.04 mmol). The flask was sealed and the mixture stirred at 80 C overnight. The reaction mixture was concentrated under reduced pressure and the residue was dissolved in methanol (5 mL) and applied to an SCX cartridge (5 g). The cartridge was washed with methanol (25 mL) then eluted with 2 M ammonia in methanol (15 mL). The ammonia eluate was concentrated under vacuum to give the desired compound (172 mg, 98%); ¹ H NMR (300 MHz, CDC ) 6.99 (m, 1 H), 6.82 - 6.74 (m, 2H), 5.24 (m, 1 H), 3.73 (m, 1 H), 3.32 (m, 1 H), 3.12 (m, 1 H), 2.95 - 2.75 (m, 7H), 2.64 (m, 1 H), 2.54 (m, 1 H), 2.04 (m, 1 H), 1 .57 (m, 1 H), 1 .43 (s, 9H).

(b) 1 -Amino-3-((6-fluoro- 1 ,2,3,4-tetrahydronaphthalen-2-yl)amino)propan-2-ol (A20) f erf-Butyl (3-((6-fluoro-1.2,3,4-tetrahydronaphthalen-2-yl)amino)-2-hydroxypropyl)carbamate

A19 (172 mg, 0.51 mmol) was dissolved in DCM (7 mL) and to the solution was added trifluoroacetic acid (697 uL, 10.2 mmol). The reaction was stirred at room temperature under a nitrogen atmosphere for 22 hours and the solvent was removed under reduced pressure. The resulting oil was dissolved in methanol (5 mL) and applied to an SCX cartridge. The cartridge was washed with methanol (25 mL) and eluted with 2 M ammonia in ethanol (15 mL). The ammonia eluate was concentrated under vacuum to give the desired compound (121 mg, 100 %); ¹H NMR (300 MHz, CDC ) 6.98 (m, 1 H), 6.78 (m, 2H), 3.65 (m, 1 H), 3.01 - 2.48 (m, 13H), 2.01 (m, 1 H), 1.57 (m, 1 H).

(c) 4-Chloro-N -(3-((6-fluoro-1 ,2,3,4-tetrahydronaphthalen-2-yl)amino)-2- hydroxypropyl)benzamide (32)

To a solution of 4-chlorobenzoic acid (88 mg, 0.56 mmol) in DCM (7.0 mL) was added N,N- diisopropylethylamine (294 pL, 1.69 mmol), A/-ethyl-A/'-(3-dimethylaminopropyl)carbodiimide hydrochloride (140.6 mg, 0.73 mmol) and 1 -hydroxybenzotriazole hydrate (1 12 mg, 0.73 mmol) under a nitrogen atmosphere. The solution was stirred for 20 minutes at room temperature and 1-amino-3-((6-fluoro-1 ,2.3,4-tetrahydronaphthalen-2-yl)amino)propan-2-ol A20 (121 mg, 0.51 mmol) was added. The mixture was stirred at room temperature for 24 hours. Water (25 mL) and DCM (25 mL) were added and the aqueous phase was extracted with DCM (3 x 20 mL). The combined organic extracts were washed with water (50 mL), brine (50 mL), dried over sodium sulfate and concentrated. The resulting oil was dissolved in methanol (5 mL) and applied to an SCX cartridge (5 g). The cartridge was washed with methanol (30 mL) then eluted with 2 M ammonia in ethanol (20 mL). The ammonia eluate was concentrated and the resulting residue was purified by preparative HPLC (Waters) to give the desired compound (2 mg, 1 %); LCMS-D RT 5.07 min; m/z 377.2.

Example 18: 4-Chloro-/V-{2-hydroxy-3-(methyl(1 ,2,3,4-tetrahydronaphthalen-2- yl)amino)propyl) benzamide (33)

(a) 2-(2-Hydroxy-3-(methyl( 1.2, 3,4-tetrahydronaphthalen-2-yl)amino)propyl)isoindoline- 1,3- dione (A21)

A mixture of A/-(2,3-epoxypropyl)phthalimide (0.463 g, 2.27 mmol) and /V-methyl-1 ,2.3,4- tetrahydronaphthalen-2-amine (0.368 ml_, 2.17 mmol) in ethanol (3 mL) was refluxed for 4 hours under a nitrogen atmosphere. The volatiles were removed in vacuo. The material was adsorbed onto silica gel and purified by column chromatography (12 g S1O2 Cartridge, 0- 15% methanol in dichloromethane) to give the desired compound as a light yellow coloured oil (0.525 g, 66%). H NMR (300 MHz, CDCI3) δ 7.89 - 7.85 (m, 2H), 7.75 - 7.72 (m, 2H), 7.14 - 7.07 (m, 4H), 4.21 - 4.09 (br. s., 1 H), 3.90 - 3.73 (m, 2H), 3.04 - 2.72 (m, 8H), 2.49 (br. s., 3H), 2.19 - 2.06 (m, 1 H), 1 .80 - 1.66 (m, 1 H).

(b) 1 -Amino-3-(methyl( 1,2,3, 4-tetrahydronaphthalen-2-yl)amino)propan-2-ol (A22)

A mixture of 2-(2-hydroxy-3-(methyl(1 ,2,3,4-tetrahydronaphthalen-2- yl)amino)propyl)isoindoline-1 ,3-dione A21 (0.525 g, 1 .44 mmol) and hydrazine monohydrate (0.143 mL, 2.88 mmol) in ethanol (2 mL) and dichloromethane (2 mL) was stirred at room temperature for 20 hours under a nitrogen atmosphere. The volatiles were removed by rotary evaporation. The residue was dissolved in methanol and passed through a SCX cartridge (5g, Agilent, Bond Elut, Mega BE-SCX) washing with 100% methanol and eluting with 100% 2 M ammonia in methanol solution to give the desired compound (0.283 g, 83%). The material was used in the next step without further purification. (c) 4-Chloro-N-(2-hydroxy-3-(methyl(1,2,3,4-tetrahydronaphthalen-2- yl)amino)propyl)benzamide (33)

A solution of 4-chlorobenzoic acid (0.208 g, 1.32 mmol) in Λ, V-dimethylformamide (3 mL) containing A/./V-diisopropylethylamine (0.841 mL, 4.83 mmol), /V-ethyl- V-(3- dimethylaminopropyl)carbodiimide hydrochloride (0.347 g, 1.81 mmol) and 1 - hydroxybenzotriazole hydrate (wetted with not less than 20 wt. % water, 0.277 g, 1.81 mmol) was stirred at room temperature for 20 minutes. After this time, a solution of 1 -amino-3- (methyl(1.2,3,4-tetrahydronaphthalen-2-yl)amino)propan-2-ol A22 (0.283 g, 1.20 mmol) in A/,/V-dimethylformamide (3 mL) was added and stirred at room temperature for 20 hours under a nitrogen atmosphere. Water and ethyl acetate were added. The aqueous phase was extracted with ethyl acetate (2 20 mL) and the combined organic extracts were washed with water (2 20 mL), brine, dried (sodium sulfate), filtered and concentrated under reduced pressure. The material was purified by column chromatography (12 g S1O2

Cartridge, 0-10% methanol in dichloromethane) to give the desired compound as a white solid (0.279 g, 61 %). H NMR (300 MHz, c₄-methanol) δ 7.81 (dd, J=8.52, 1 .41 Hz, 2H), 7.48 (dd, J=8.58, 0.99 Hz. 2H), 7.05 - 7.04 (m, 4H), 3.99 - 3.93 (m, 1 H), 3.54 (dd, J=13.53, 4.89 Hz, 1 H), 3.44 - 3.35 (m, 1 H), 2.92 - 2.80 (m, 5H), 2.68 - 2.63 (m, 2H), 2.43 (s, 3H), 2.10 - 2.02 (m, 1 H), 1.70 - 1.59 (m, 1 H). LCMS-D: RT 5.12 min; m/z 373.3 [M+H]⁺.

Example 19: A -{3-{Bicyclo[4.2.0]octa-1(6),2,4-trien-7-ylamino)-2-hydroxypropyl)-4- chlorobenzamide 34)

A10 34

A mixture of A -(3-amino-2-hydroxypropyl)-4-chlorobenzamide hydrochloride A10 (0.080 g, 0.30 mmol), A/,/V-diisopropylethylamine (0.053 mL, 0.30 mmol), benzocylclobuten-1 (2H)-one (0.039 mL, 0.33 mmol) and titanium(IV) isopropoxide (0.1 19 mL, 0.392 mmol) in absolute ethanol (2 mL) was stirred at 30 °C for 16 hours under a nitrogen atmosphere. Sodium borohydride (0.023 g, 0.60 mmol) was added and the mixture was stirred for 6 hours under a nitrogen atmosphere. The reaction mixture was quenched with a saturated aqueous sodium hydrogen carbonate solution and ethyl acetate. The precipitated inorganic solid was separated by filtration and the aqueous layer was extracted with ethyl acetate (2 15 mL). The combined organic extracts were washed with water, brine, dried (sodium sulfate), filtered and concentrated under reduced pressure. The material was purified by column chromatography (12 g S1O2 cartridge, 0-15% methanol in DCM) to give the desired compound as a white solid (0.028 g, 28%). ¹H NMR (300 MHz, d_t-methanol) δ 7.80 (d, J=8.61 Hz, 2H), 7.46 (d, J=8.61 Hz, 2H), 7.30 (t, J=3.85 Hz, 1 H), 7.22 - 7.1 1 (m, 3H), 4.05 - 3.94 (m. 1 H), 3.91 - 3.77 (m, 2H), 3.51 - 3.43 (m, 1 H), 2.88 - 2.65 (m, 2H), 2.37 (s, 2H). LCMS-D: RT 4.77 min; m/z 331 .2 [M+H]\

Example 20: 2-((1 -Acetylpiperidin-4-yl)amino)-W-{3-{(2,3-dihydro-1 H-inden-2-yl)amino)- 2-h droxypropyl)isonicotinamide (35)

35

(a) Methyl 2-(( 1 -acetylpiperidin-4-yl)amino)isonicotinate (A23)

A mixture of 1 -(4-aminopiperidin-1-yl)ethan-1-one hydrochloride (0.500 g, 2.80 mmol), methyl 2-bromoisonicotinate (0.423 g, 1 .96 mmol), CS2CO3 (2.55 g, 7.84 mmol), xantphos (0.057 g, 0.098 mmol) and Pd₂(dba)₃ (0.090 g, 0.098 mmol) in 1 ,4-dioxane (15 mL) was bubbled with nitrogen for 10 minutes. The mixture was stirred under an atmosphere of nitrogen at 80 C for 16 hours. Another portion of CS2CO3 (2.55 g, 7.84 mmol), xantphos (0.057 g, 0.098 mmol) and Pd₂(dba)₃ (0.090 g, 0.098 mmol) was added. The mixture was stirred under an atmosphere of nitrogen at 80 C for 4 hours. The reaction mixture was cooled to room temperature and diluted with EtOAc (-50 mL). Solid impurities were removed by filtration and the filtrate was removed in vacuo. The resultant solid was purified by column chromatography (40 g S1O2 cartridge, 0-100% EtOAc in petroleum benzine 40-60 C, then 0- 20% MeOH in EtOAc) to give the desired compound as a yellow oil (0.268 g, 49%). LCMS- B: RT 3.08 min; m/z 278.2 [M+H]⁺.

(b) 2-(( 1 -Acetylpiperidin-4-yl)amino)isonicotinic acid (A24)

A mixture of methyl 2-((1-acetylpiperidin-4-yl)amino)isonicotinate A23 (0.268 g, 0.966 mmol), LiOH.h O (0.811 g, 19.3 mmol), THF (7 mi_), MeOH (7 mL) and H₂0 (1.5 mL) was stirred at 40 C for 2 hours. The mixture was cooled to room temperature and the volatiles were removed. Water (-20 mL) was added and the pH was adjusted to ~6 with aqueous HCi (2 M). A yellow precipitate formed which was isolated by vacuum filtration and air dried to give the desired compound as a yellow solid (0.058 g, 23%). The aqueous filtrate was passed through an Oasis HLB 35 cc LP extraction cartridge (6 g) which was washed with 3 column volumes of water. The lipophilic component was eluted with 3 column volumes of MeOH and concentration in vacuo gave the desired compound as a yellow solid (0.122 g, 48%). Overall yield: 0.180 g, 71%. H NMR (400 MHz, d₆-DMSO) δ 8.08 (d, J = 5.2 Hz, 1H), 7.01 -6.95 (m, 1H).6.85 (dd, J = 5.2, 1.4 Hz, 1H), 6.81 (d, J = 7.5 Hz, 1H), 4.27-4.15 (m, 1H), 4.02- 3.91 (m, 1 H), 3.82 - 3.71 (m, 1 H), 3.22 - 3.11 (m, 1 H), 2.86 - 2.74 (m, 1 H), 2.00 (s, 3H), 1.97- 1.83 (m, 2H), 1.41 - 1.14 (m, 2H); LCMS-B: RT 1.26 min; m/z 264.2 [M+H]⁺

(c) 2-(( 1 -Acetylpiperidin-4-yl)amino)- -(3-((2, 3-dihydro- 1 H -inden-2-yl)amino)-2- hydroxypropyl)isonicotinamide (35)

A mixture of 2-((1 -acetylpiperidin-4-yl)amino)isonicotinic acid A24 (0.122 g, 0.463 mmol), triethylamine (258 L, 1.85 mmol), and DCM (10 mL) was cooled to 0 C under a nitrogen atmosphere and isobutyl chloroformate (61 L, 0.46 mmol) was added. The mixture was stirred for 30 minutes at 0 C before being transferred via syringe to a solution of 1-amino-3- ((2.3-dihydro-1H-inden-2-yl)amino)propan-2-ol I2 (0.096 g, 0.46 mmol) in DCM (10 mL) at 0 C After 2.5 hours, the mixture was diluted with DCM (10 mL), water (10 mL) and aqueous sodium hydroxide (2 M, 10 mL). The separated aqueous phase was extracted with DCM (2 15 mL). The combined organic extracts were washed with brine (25 mL), dried (MgSO.i) and concentrated in vacuo. The residue was purified by column chromatography (12 g Si02 cartridge, 0-35% MeOH in CHCI3) to give the desired compound as a yellow oil (0.012 g, 6%).'H NMR (400 MHz, d₆-DMSO) δ 8.76 (t, J = 5.9, 5.9 Hz, 1H).8.05 (d, J = 5.2 Hz, 1H), 7.26 - 7.22 (m, 2H), 7.20-7.16 (m, 2H), 6.94-6.87 (m, 2H), 6.76 (d, J = 7.6 Hz, 1H), 5.78 (s, 1H), 4.26-4.17 (m, 1H), 4.03-3.91 (m, 3H), 3.82-3.72 (m, 1H), 3.32-3.20 (m, 4H), 3.17-3.06 (m, 5H), 2.91 -2.83 (m, 1H), 2.83-2.74 (m, 1H), 2.00 (s, 3H), 1.95- 1.82 (m, 2H), 1.41 - 1.21 (m, 2H); LCMS-A: RT 4.06 min; m/z 452.2 [M+H]\ Example 21 : (S)-4-((3-((2,3-Dihydro-1 H-inden-2-yl)amino)-2- hydroxypropyl)carbamoyl)benzoic acid (36)

36

(a) Methyl (S)-4-((3-((2,3-dihydro- 1 H -inden-2-yl)amino)-2- hydroxypropyl)carbamoyl)benzoate (A25)

To a solution of 4-(methoxycarbonyl)benzoic acid (0.96 g, 5.3 mmol) in DCM (30 mL) at 0 C was added HATU (2.77 g, 7.28 mmol). A solution of (f?)-1 -amino-3-((2,3-dihydro-1 H-inden-2- yl)amino)propan-2-ol (1.0 g, 4.9 mmol) in DCM (20 mL) and DIPEA (1.87 mL 14.6 mmol) were added and the mixture stirred at room temperature overnight. Water (25 mL) and DCM (25 mL) were added and the organic layer separated. The aqueous layer was further extracted with DCM (2 25 mL) and the combined organic layers dried (NazSOi), filtered and concentrated in vacuo. The residue obtained was purified by column chromatography (ethyl acetate : petroleum ether = 1 :50, then DCM, then DCM : methanol 10:1 ) to give the desired compound as a white solid (958 mg, 53%). LCMS-E: RT 1 .86 min; m/z 369.2

[M+H]\

(b) (S)-4-((3-((2,3-Dihydro- 1 H -inden-2-yl)amino)-2-hydroxypropyl)carbamoyl)benzoic acid

(36)

To a solution of (S)-methyl 4-((3-((2,3-dihydro-1 H-inden-2-yl) amino)-2-hydroxypropyl) carbamoyl) benzoate A25 (935 mg, 2.54 mmol) in a mixture of THF (15 mL), methanol (8 mL) and water (4 mL) was added UOH.H2O (533 mg, 12.7 mmol). The reaction mixture was stirred at room temperature overnight. The mixture was concentrated and the remaining aqueous solution acidified to pH = 5-6 with 2 M HCI. The solid precipitate that formed was collected by filtration, washed with water and dried to give the desired compound as a white solid (610 mg, 67%). LCMS-E: RT 0.69 min; m/z 355.2 [M+H]⁺. Example 22: /V-((S)-3-((2,3-Dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)-4-((fl)-3- meth lmorpholine-4-carbonyl)benzamide (37)

To a solution of (S)-4-((3-((2,3-dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)carbamoyl) benzoic acid 36 (50 mg, 0.14 mmol) in DMF (2 mL) were added HATU (80 mg, 0.21 mmol), triethylamine ( 52 mg, 0.52 mmol) and (f?)-3-methylmorpholine (17 mg, 0.17 mmol). The resultant mixture was stirred at room temperature overnight. The mixture was diluted with water (20 mL) and extracted with DCM (20 mL 3). The combined organic layers were washed with water (30 mL 2) and brine (30 mL 4), dried ( a2S04), filtered and concentrated under reduced pressure. The residue obtained was purified by Preparative TLC (DCM : MeOH = 10: 1 ) to give the desired product as a yellow solid (14 mg, 23%). LCMS-C: RT 0.78 min; m/z 438.3 [M+H]\ Example 23: A -((S)-3-((2,3-Dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)-4-{(S)-3- meth lmorpholine-4-carbonyl)benzamide (38)

To a solution of (S)-4-((3-((2,3-dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)carbamoyl) benzoic acid 36 (50 mg, 0.14 mmol) in DMF (2 mL) were added HATU (80 mg, 0.21 mmol), triethylamine (52 mg, 0.52 mmol) and (S)-3-methylmorpholine (17 mg, 0.17 mmol). The resultant mixture was stirred at room temperature overnight. The mixture was diluted with water (20 mL) and extracted with DCM (20 mL 3). The combined organic layers were washed with brine (30 mL χ 3), dried (Na2S0₄), filtered and concentrated under reduced pressure. The residue was purified by Preparative TLC (DCM:MeOH = 10:1 ) to give the desired compound as a white solid (6 mg, 10%); LCMS-C: RT 0.91 min; m/z 438.3 [M+H] \ Example 24: 4-(8-Oxa-3-azabicyclo[3.2.1]octane-3-carbonyl)-N-(3-((2,3-dihydro-1 H- inden-2- l)amino)-2-hydroxypropyl)benzamide (39)

19 39 To a solution of 4-((3-((2,3-dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)carbamoyl) benzoic acid 19 (50 mg, 0.14 mmol) in a mixture of DCM (1 5 mL) and DMF (5 mL) were added HATU (76 mg, 0.20 mmol) and DIPEA (82 mg, 0.63 mmol). The mixture was stirred at room temperature for 30 minutes, then 8-oxa-3-azabicyclo[3.2.1 ]octane hydrochloride (25.4 mg, 0.17 mmol) was added. The resulting mixture was stirred at room temperature overnight. The mixture was diluted with water (20 mL) and extracted with DCM (30 mL 4). The combined organic layers were washed with brine (30 mL), dried (NazSC ), filtered and concentrated under reduced pressure. The residue was purified by Preparative-TLC (DCM :MeOH = 10: 1 ) to give the desired product as an off-white solid (20 mg, 32%). LCMS-C: RT 0.83 min; m/z 450.2 [M+H]\

Example 25: 4-(3-Oxa-8-azabicyclo[3.2.1]octane-8-carbonyl)-A -(3-((2,3-dihyd

ind -2-yl)amino)-2-hydroxypropyl)benzamide (40)

To a solution of 4-((3-((2,3-dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)carbamoyl) benzoic acid 19 (50 mg, 0.14 mmol) in DCM (4 mL) were added HATU (80 mg, 0.21 mmol) and DI PEA (73 mg, 0.56 mmol). The mixture was stirred at room temperature for 30 minutes. 3-Oxa-8-azabicyclo[3.2.1 ]octane hydrochloride (25 mg, 0.17 mmol) was added and the resulting mixture was stirred at room temperature overnight. The mixture was diluted with water (20 mL) and extracted with DCM (10 mL ^χ 5). The combined organic layers were washed with brine (10 mL 3), dried ( a2SC>4), filtered and concentrated under reduced pressure. The residue was purified by Preparative TLC (DCM:MeOH = 15:1 ) to give the desired compound as an off-white solid (35 mg, 56%). LCMS-C: RT 1 .03 min; m/z 450.3 [M+H]\ Example 26: i¥-(3-((2,3-Dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)-4-(2,2- dimeth lmorpholine-4-carbonyS)benzamide (41 )

19 41

To a solution of 4-((3-((2,3-dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)carbamoyl) benzoic acid 19 (50 mg, 0.14 mmoi) in a mixture of DCM (15 mL) and DMF (5 ml_) were added HATU (76 mg, 0.20 mmoi) and DIPEA (82 mg, 0.63 mmoi). The mixture was stirred at room temperature for 30 minutes, then 2.2-dimethylmorpholine (19.6 mg, 0.17 mmoi) was added and the resulting mixture stirred at room temperature overnight. The mixture was diluted with water (20 mL) and extracted with DCM (30 mL 4). The combined organic layers were washed with water (30 mL 3) and brine (30 mL), dried filtered and concentrated under reduced pressure. The residue was purified by Preparative TLC (DCM : MeOH = 10: 1 ) to give the desired compound as an off-white solid (15 mg, 24%).LCMS-C: RT 1 .24 min; m/z 452.3 [M+H]\

Example 27: (S)-3-((1 -Acetylpiperidin-4-yl)amino)-/V-(3-((2,3-dihydro-1 H-inden-2- l)amino)-2-hydroxypropyl)benzamide (42)

(a) tert-Butyl 4-((3-(methoxycarbonyl)phenyl)amino)piperidine- 1 -carboxylate A26

A mixture of 1 -Boc-4-piperidone (0.725 g, 3.64 mmoi), methyl 3-aminobenzoate (0.500 g, 3.31 mmoi) and acetic acid (0.38 mL, 6.6 mmoi) in DCE (25 mL) was stirred at room temperature for 15 minutes before sodium triacetoxyborohydride (1.05 g, 4.96 mmoi) was added. The mixture was stirred for a further 16 hours at room temperature and then quenched by the addition of H₂0 (20 mL). The mixture was concentrated in vacuo, saturated aqueous NaHCCh (50 mL) was added and the aqueous was extracted with EtOAc (3 * 75 mL). The organic layers were combined, washed with brine, dried (MgS0₄) and the solvent removed in vacuo. The solid residue was purified by column chromatography (Biotage Isolera, 40 g S1O2 cartridge, 0-30% EtOAc in petroleum benzine 40-60 C) to give the desired compound as a colourless oil (1 .35 g, -80% purity, >95% yield). ¹ H NMR (400 MHz, chloroform- ) δ 7.46 - 7.40 (m, 1 H), 7.38 - 7.32 (m, 1 H), 7.25 - 7.17 (m, 1 H), 6.92 - 6.84 (m, 1 H), 4.07 - 3.99 (m, 1 H), 3.90 - 3.86 (m, 3H), 3.71 (t, J = 6.2 Hz, 1 H), 3.47 (it J = 10.4, 3.9 Hz, 1 H), 2.90 (t, J = 12.5 Hz, 2H), 2.43 (t, J = 6.2 Hz, 1 H), 2.10 - 1.97 (m, 2H), 1 .46 (s. 9H), 1 .41 - 1 .31 (m, 2H). LCMS-A: RT 6.70 min; m/z 279.1 [M-t-Bu+2H]\

(b) 3-((1-(tert-Butoxycarbonyl)piperidin-4-yl)amino)benzoic acid A27

A mixture of ie/t-butyl 4-((3-(methoxycarbonyl)phenyl)amino)piperidine-1 -carboxylate A26 (1 .35 g, -80 purity, 3.23 mmoi), LiOH.H₂0 (1 .69 g, 40.4 mmoi), THF (14 mL), MeOH (14 mL) and H2O (3 mL) was stirred overnight at room temperature. The reaction mixture was concentrated in vacuo, H2O (50 mL) and aqueous NaOH (2 M, 30 mL) were added and the aqueous solution was washed with EtOAc (3 * 50 mL). The aqueous phase was acidified with an aqueous HCI solution (2 M) and extracted with EtOAc (3 * 50 mL). The organic layers were combined, washed with brine, dried (MgSO.4) and the solvent removed in vacuo to give the desired compound as a yellow oil (0.801 g, 77%). Ή NMR (400 MHz, DMSO-cfe) 5 7.20 - 7.14 (m, 2H), 7.12 - 7.08 (m, 1 H), 6.84 - 6.77 (m, 1 H), 5.78 (s, 1 H), 3.92 - 3.80 (m, 2H), 3.49 - 3.39 (m. 1 H). 3.02 - 2.82 (m, 2H), 1 .89 - 1.82 (m, 2H), 1.40 (s, 9H), 1.28 - 1.19 (m, 2H). LCMS-A: RT 6.04 min; m/z 265.1 [M-t-Bu+2H]+; 319.1 [M-H]\

(c) tert-Butyl (R)-4-((3-((3-((2,3-dihydro- 1H-inden-2-yl)(2A-dimethoxybenzyl)am ^ hydroxypropyl)carbamoyl)phenyl)amino)piperidine- 1 -carboxylate A28

A solution of 3-((1-(fe/?-butoxycarbonyl)piperidin-4-yl)amino)benzoic acid A27 (0.099 g, 0.31 mmoi) and DIPEA (98 μΙ_, 0.56 mmoi) in DCM (5 mL) was stirred with HATU (0.1 17 g, 0.309 mmoi) for 10 minutes at room temperature. A solution of (f?)-1 -amino-3-((2.3-dihydro-1 H- inden-2-yl)(2,4-dimethoxybenzyl)amino)propan-2-ol 110 (0.100 g, 0.281 mmoi) in DCM (5 mL) was then added dropwise and the mixture was stirred at room temperature for 24 h. Water (30 mL) and aqueous NaOH (2 M, 10 mL) were then added and the aqueous was extracted with DCM (3 * 30 mL). The organic layers were combined, the solvent was removed in vacuo and the residue was purified by column chromatography (Biotage Isolera, 24 g Si0₂ cartridge, 0-100% EtOAc in petroleum benzine 40-60 C) to give the desired compound as a colourless oil (0.105 g, 56%). ¹ H NMR (400 MHz, chloroform-d) δ 7.18 - 7.08 (m, 6H), 7.01 (t, J = 2.0 Hz, 1 H), 6.88 (d, J = 7.6 Hz, 1 H), 6.71 - 6.60 (m, 2H), 6.43 (d, J = 2.4 Hz, 1 H), 6.39 (dd, J = 8.3, 2.4 Hz, 1 H), 4.07 - 3.98 (m, 2H), 3.87 - 3.81 (m, 2H), 3.79 - 3.73 (m, 7H), 3.71 - 3.54 (m, 3H), 3.51 - 3.40 (m, 1 H), 3.36 - 3.24 (m, 1 H). 3.14 - 2.98 (m, 4H), 2.95 - 2.86 (m, 2H), 2.65 - 2.48 (m, 2H), 2.01 - 1.97 (m, 1 H), 1.46 (s, 9H), 1.36 - 1.28 (m, 2H); N-H and O-H not observed. LCMS-A: RT 5.20 min; m/z 659.3 [M+H]\

(d) (R)-N-(3-((2.3-Dihydro-1H-inden-2-yl)(2,4-dimethoxybenzyl)amino)-2-hydroxypropyl)-3- (piperidin-4-ylamino jbenzamide A29

A mixture of ieri-butyl (f?)-4-((3-((3-((2,3-dihydro-1 H-inden-2-yl)(2,4- dimethoxybenzyl)amino)-2-hydroxypropyl)carbamoyl)phenyl)amino)piperidine-1 -carboxylate A28 (0.105 g, 0.159 mmol) and TFA (122 μΙ_, 1.59 mmol) in DCM (10 mL) was stirred at room temperature for 16 hours. Water (20 mL) was added and the aqueous was washed with DCM (3 x 10 mL). The aqueous was adjusted to a pH of -14 with an aqueous NaOH solution (2 M) and then extracted with DCM (3 x 15 mL). The organic layers were combined, washed with brine, dried (MgSCu) and the solvent removed in vacuo to give the desired compound as a white powder (0.024 g, 27%). Ή NMR (400 MHz, DMSO-cfe) δ 8.08 (t, J = 5.4 Hz, 1 H), 7.28 (d, J = 8.3 Hz, 1 H), 7.19 - 7.04 (m, 5H), 6.96 (s, 1 H), 6.88 (d, J = 7.6 Hz, 1 H), 6.69 (dd, J = 8.1 , 2.3 Hz, 1 H), 6.48 (d, J = 2.4 Hz, 1 H), 6.42 (dd, J = 8.3, 2.4 Hz, 1 H), 5.59 (d, J = 8.1 Hz, 1 H), 4.66 (d, J = 4.3 Hz, 1 H), 3.83 - 3.75 (m, 1 H), 3.73 - 3.68 (m, 7H), 3.57 (d, J = 2.8 Hz, 2H), 3.27 - 3.09 (m, obscured by solvent), 3.01 - 2.80 (m, 7H), 2.47 - 2.39 (m, obscured by solvent), 1.88 - 1.77 (m, 2H), 1.26 - 1.14 (m, 3H). (e) (R)-3-((1-Acetylpiperidin-4-yl)amino)-N-(3-((2,3-dihydm

dimethoxybenzyl)amino)-2-hydroxypropyl)benzamide A30

A solution of (f?)-/V-(3-((2,3-dihydro-1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino)-2- hydroxypropyl)-3-(piperidin-4-ylamino)benzamide A29 (0.024 g, 0.043 mmol) and

triethylamine (12 μί, 0.086 mmol) in DCM (4 mL) was cooled to 0 °C before acetic anhydride (4 L, 0.05 mmol) was added. The mixture was stirred and allowed to return to room temperature over 3 hours. Water (10 mL) and an aqueous solution of NaOH (2 M, 10 mL) were added and the mixture was stirred for 20 minutes. The aqueous was extracted with DCM (3 15 mL), the organic layers were combined and the solvent removed in vacuo to give the desired compound as a white solid (0.026 g, >95%). LCMS-B: RT 3.38 min; m/z 601.3 [M+H]\ (f) (S)-3-((1 -Acetylpiperidin-4-yl)amino)-N-(3-((2,3-dihydro- 1 H-inden-2-yl)amino)-2- hydroxypropyl)benzamide 42

A mixture of (f?)-3-((1-acetylpiperidin-4-yi)amino)-/V-(3-((2,3-dihydro-1 H-inden-2-yl)(2,4- dimethoxybenzyl)amino)-2-hydroxypropyl)benzamide A30 (0.026 g, 0.043 mmol) and TFA (2 mL) was stirred at 70 C for 16 hours. The mixture was returned to room temperature, water was added (20 mL) and the aqueous was washed with DCM (3 ^χ 20 mL). The pH of the aqueous layer was adjusted to -14 with an aqueous NaOH solution (2 M) and then extracted with DCM (3 x 20 mL). The organic layers were combined and the solvent removed in vacuo to give the desired compound as a white solid (0.017 g, 87%). ¹H NMR (400 MHz, DMSO-cfe) 5 8.26 (t. J = 5.7 Hz, 1 H), 7.21 - 7.03 (m, 6H), 6.94 (d, J = 7.6 Hz, 1 H), 6.78 - 6.66 (m, 1 H), 5.70 (d, J = 8.2 Hz, 1 H), 4.97 - 4.76 (m, 1 H), 4.29 - 4.14 (m. 1 H), 3.83 - 3.73 (m, 1 H), 3.72 - 3.65 (m, 1 H), 3.54 - 3.45 (m, 2H), 3.30 - 3.21 (m, 2H). 3.21 - 3.13 (m, 1 H), 3.09 - 2.99 (m, 2H), 2.86 - 2.75 (m, 1 H), 2.70 - 2.50 (m, 4H), 2.00 (s, 3H), 1 .96 - 1.82 (m, 2H), 1 .36 - 1.12 (m, 2H); N-H not observed. LCMS-B:RT 3.20 min; m/z 451 .3 [M+H]\

Exam le 28: Amide couplings

Method X1 - A solution of HATU (0.080 g, 0.21 mmol, 1 .5 equiv) in CH₃CN (1 mL) was added to the desired acid (0.14 mmol, 1 equiv.), CH₃CN (1 mL) and DIPEA (73 pL, 0.42 mmol, 3 equiv.) and the mixture was stirred for 10 minutes. A solution of 1 -amino-3-((2,3- dihydro-1 /-/-inden-2-yl)(2,4-dimethoxybenzyl)amino)propan-2-ol I2 (0.050 g, 0.14 mmol, 1 equiv) in DMF (1 mL) was then added and the mixture was stirred at room temperature for 16 h. DCM (1 mL) and a solution of NaOH (aqueous 2 M, 2 mL) were added and the phases were separated. The aqueous phase was extracted with DCM (2 mL) and the organic layers were combined. The organic extracts were concentrated by a stream of air and the resultant residue dissolved in MeOH and purified by solid-phase extraction (1 g SCX-2 cartridge, 4 column volumes of methanol followed by 4 column volumes of 7 N NH₃ in MeOH). The basic fractions were concentrated by a stream of air, taken up in TFA (0.5 mL) and stirred at 70 C for 5 hours. Water (2 mL) was added and the aqueous solution was washed with DCM (3 * 2 mL). The aqueous layer was brought to ~pH 14 by the addition of an aqueous NaOH solution (2 M) and then extracted with DCM (2 2 mL). The organic layers were

concentrated with a stream of air to give the desired compounds.

Example 29: 2-((1 -Acetylpiperidin-4-yl)oxy)-A -(3-{(2,3-dihydro-1 H-inden-2-yl)amino)-2- h droxypropyl)isonicotinamide 50

(a) 2-((1-(tert-Butoxycarbonyl)piperidin-4-yl)oxy)isonicotinic acid A31

A solution of ie t-butyl 4-hydroxypiperidine-1-carboxylate (0.500 g, 2.48 mmol) in anhydrous DMF (5 mL) was added to a stirring suspension of sodium hydride (60% dispersion in mineral oil; 0.238 g, 9.94 mmol) in anhydrous DMF (5 mL) under an atmosphere of nitrogen. The mixture was stirred at room temperature for 10 minutes before a solution of 2- fluoroisonicotinic acid (0.319 g, 2.26 mmol) in DMF (5 mL) was added. The mixture was stirred for a further 16 hours at room temperature and 4 hours at 60 °C. After returning to room temperature, H₂0 (20 mL) was carefully added and the pH of the aqueous mixture was adjusted to ~2 with an aqueous HCI solution (2 M). The aqueous solution was extracted with EtOAc (3 x 30 mL), the organic layers were combined, washed with brine, dried (MgS0₄) and the solvent removed in vacuo. The resultant residue was purified by column

chromatography (Biotage I solera, 40 g S1O2 cartridge, 0-100% EtOAc in petroleum benzine 40-60 C) to give a white solid that was an approximate 1 :1 mixture of the desired product and the isonicotinic acid starting material. This material was dissolved in DMF (5 mL) and added to a mixture of iert-butyl 4-hydroxypiperidine-1-carboxylate (0.500 g, 2.48 mmol) in anhydrous DMF (5 mL) that had been added to a stirring suspension of sodium hydride (60% dispersion in mineral oil; 0.238 g, 9.94 mmol) in anhydrous DMF (5 mL) under an atmosphere of nitrogen. The mixture was stirred for 16 hours before H2O (20 mL) was carefully added and the pH of the aqueous mixture was adjusted to ~2 with an aqueous HCI solution (2 M). The aqueous solution was extracted with DCM (3 * 30 mL), the organic layers were combined, dried (MgS0₄) and the solvent removed in vacuo. The resultant residue was purified by column chromatography (Biotage Isolera, 40 g S1O2 cartridge, 0- 100% EtOAc in petroleum benzine 40-60 °C) to give the desired compound as a white solid (0.419 g, 58%). ¹H NM (400 MHz, DMSO-ofe) δ 8.34 - 8.27 (m, 1 H), 7.39 - 7.34 (m, 1 H), 7.17 - 7.13 (m, 1 H), 5.27 - 5.15 (m, 1 H), 3.74 - 3.62 (m, 2H), 3.23 - 3.1 1 (m, 2H), 2.00 - 1.88 (m, 2H), 1.63 - 1.49 (m, 2H), 1.40 (s, 9H); COOH not observed. LCMS-B: RT 3.65 min; m/z 321 .2 [M-H]-.

(b) tert-Butyl 4-((4-((3-((2,3-dihydro- 1H-inden-2-yl)(2,4-dimethoxybenzyl)amino)-2- hydroxypropyl)carbamoyl)pyridin-2-yl)oxy)piperidine- 1 -carboxylate A32

A mixture of 2-((1 -(iert-butoxycarbonyl)piperidin-4-yl)oxy)isonicotinic acid A31 (0.100 g, 0.310 mmol), DIPEA (108 μΙ_, 0.620 mmol) and HATU (0.1 18 g, 0.310 mmol) in DCM (5 mL) was stirred for 15 minutes at room temperature under an atmosphere of nitrogen. A solution of 1-amino-3-((2.3-dihydro-1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino)propan-2-ol 112 (0.1 1 1 g, 0.310 mmol) in DCM (5 mL) was then added and the mixture was stirred for 16 hours at room temperature. An aqueous NaOH solution (2 M, 20 mL) was added and the aqueous was extracted with DCM (3 * 30 mL). The organic layers were combined, dried (MgSC ), filtered, and the volatiles removed in vacuo. The residue was purified by column chromatography (Biotage Isolera, 24 g S1O2 cartridge, 0-10% MeOH in DCM) to give the desired compound as a brown oil (0.204 g, >95%). ¹H NMR (400 MHz, DMSO-cfe) δ 8.53 (t, J = 5.6 Hz, 1 H), 8.24 (d, J = 5.2 Hz, 1 H), 7.31 - 7.24 (m, 2H), 7.19 - 7.04 (m, 5H), 6.48 (d, J = 2.4 Hz, 1 H), 6.42 (dd, J = 8.3, 2.4 Hz, 1 H), 5.26 - 5.14 (m. 1 H), 4.67 (d, J = 4.3 Hz, 1 H), 3.85 - 3.76 (m, 1 H), 3.76 - 3.63 (m, 10H), 3.57 (s, 2H), 3.25 - 3.09 (m, 3H), 3.01 - 2.78 (m, 4H), 2.45 (d, J = 6.0 Hz, 2H), 2.00 - 1.89 (m, 2H), 1 .62 - 1.51 (m, 2H), 1 .41 (s, 9H). LCMS- A: RT 2.10 min; m/z 661 .3 [M+H]\

(c) N-(3-((2, 3-Dihydro- 1 H-inden-2-yl)(2A-dimethoxybenzyl)amino)-2-hydroxypropyl) -2- (piperidin-4-yloxy)isonicotinamide A33

A mixture of ieri-butyl 4-((4-((3-((2,3-dihydro-1 H-inden-2-yl)(2.4-dimethoxybenzyl)amino)-2- hydroxypropyl)carbamoyl)pyridin-2-yl)oxy)piperidine-1-carboxylate A32 (0.184 g, 0.278 mmol), TFA (0.32 mL, 4.2 mmol) and DCM (10 mL) was stirred at room temperature for 1 hour. Another aliquot of TFA (0.1 1 mL, 1.4 mmol) was added and the mixture was stirred at room temperature for 2.5 hours. Water (10 mL) and aqueous NaOH (2 M, 10 mL) were added and the aqueous was extracted with DCM (3 x 15 mL). The organic layers were combined and the solvent removed in vacuo to give a yellow oil that was dissolved in a minimum amount of MeOH and loaded onto an SCX cartridge (SiliCycle SCX-2 cartridge, 5 g). The cartridge was washed with 5 column volumes of MeOH before the desired component was eluted with 5 column volumes of NH3 in MeOH (3.5 M). The fractions containing the suspected product were combined and the solvent removed in vacuo to give a colourless oil. The oil was taken up in a minimum amount of DCM and cyclohexane was added to form a white precipitate. The volatiles were removed in vacuo to give the desired compound as a white solid (0.1 15 g. 74%). ¹H NMR (400 MHz, chloroform-d) δ 8.18 (d, J = 5.3 Hz, 1 H), 7.19 - 7.06 (m, 5H), 7.04 (dd, J = 5.2, 1 .5 Hz, 1 H), 7.01 - 6.95 (m, 1 H), 6.70 (t, J = 5.5 Hz, 1 H), 6.44 (d, J = 2.4 Hz, 1 H), 6.38 (dd, J = 8.2, 2.4 Hz, 1 H), 5.31 - 5.05 (m, 1 H), 3.84 - 3.76 (m, 8H), 3.62 - 3.51 (m, 2H), 3.47 (s, 2H), 3.33 - 3.24 (m, 1 H), 3.22 - 3.14 (m, 2H), 3.12 - 3.03 (m, 1 H), 3.01 - 2.83 (m, 5H), 2.53 (dd, J = 13.2, 4.1 Hz, 1 H), 2.49 - 2.41 (m, 1 H), 2.15 - 2.05 (m, 2H), 1 .80 - 1 .70 (m, 2H); N-H not observed. LCMS-A: RT 4.42 min; m/z 561 .3 [M+H]\

(d) 2-(( 1 -Acetylpiperidin-4-yl)oxy)-N-(3-((2.3-dihydro- 1 H-inden-2-yl)(2, 4- dimethoxybenzyl)amino)-2-hydroxypropyl)isonicotinamide A34

A solution of V-(3-((2,3-dihydro-1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino)-2- hydroxypropyl)-2-(piperidin-4-yloxy)isonicotinamide A33 (0.030 g, 0.054 mmol) and triethylamine (1 1 L, 0.080 mmol) in DCM (2 mL) was cooled to 0 C under an atmosphere of nitrogen. Acetic anhydride (5 L, 0.05 mmol) was added and the mixture was returned to room temperature, with stirring, over 5 hours. Water (10 mL) and an aqueous NaOH solution (2 M, 10 mL) were added and the mixture was stirred for 20 minutes. The aqueous was extracted with DCM (3x 15 mL), the organic layers were combined and the solvent removed in vacuo to give the desired compound as a yellow oil (0.032 g, >95%). LCMS-A: RT 4.79 min; m/z 603.3 [M+H]\ (e) 2-((1-Acetylpiperidin-4-yl)oxy)-N-(3-((2,3-dihydro-1H-ind

hydroxypropyl)isonicotinamide 50

A solution of 2-((1-acetylpiperidin-4-yl)oxy)-/V-(3-((2,3-dihydro-1 H-inden-2-yl)(2.4- dimethoxybenzyl)amino)-2-hydroxypropyl)isonicotinamide A34 (0.032 g, 0.053 mmol) in TFA (2 mL) was stirred at 70 °C for 4 hours. Water (25 mL) was added and the acidic aqueous phase was washed with DCM (3 15 mL). The aqueous was adjusted to pH -14 with aqueous NaOH (2 M) and extracted with DCM (3 ^χ 20 mL). The organic layers were combined and the solvent removed in vacuo to give the desired compound as a white solid (0.004 g, 15%).¹H NMR (400 MHz, DMSO-cfe) δ 8.70 (t, J= 5.7 Hz, 1H), 8.25 (d, J= 5.2 Hz, 1H), 7.33-7.27 (m, 1H), 7.21 -7.14 (m, 3H), 7.12-7.05 (m, 2H), 5.30-5.19 (m, 1H), 4.87 (s, 1H), 3.92-3.81 (m, 1H), 3.76-3.65 (m, 2H), 3.50 (m, 1H), 3.30-3.20 (m, peaks obscured by solvent), 3.11 - 2.99 (m, 2H), 2.72 - 2.61 (m, 3H), 2.55 (dd, J = 11.8, 6.8 Hz, 1H), 2.06- 1.88 (m, 5H), 1.72- 1.47 (m, 2H). LCMS-B: RT3.18 min; m/z 453.3 [M+H]\

Example 30: W-(3-((2,3-dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)-2-(piperidin-4- loxy)isonicotinamide 51

A solution of A/-(3-((2,3-dihydro-1H-inden-2-yl)(2,4-dimethoxybenzyl)amino)-2- hydroxypropyl)-2-(piperidin-4-yloxy)isonicotinamide A33 (0.020 g, 0.036 mmol) in TFA (2 mL) was stirred at 70 °C for 16 hours. The solution was concentrated in vacuo before H2O (15 mL) was added and the acidic aqueous phase was washed with DCM (3 15 mL). The aqueous phase was adjusted to pH -14 with an aqueous NaOH solution (2 M) and extracted with DCM (3 15 mL). The organic layers were combined and the solvent removed in vacuo to give the desired compound as a yellow oil (0.005 g, 34%). Ή NMR (400 MHz, DMSO-cfe) δ 8.69 (t, J = 5.8 Hz, 1 H), 8.23 (d, J = 5.5 Hz, 1 H), 7.31 - 7.24 (m, 1 H), 7.20 -7.13 (m, 3H), 7.12-7.06 (m, 2H), 5.15-5.04 (m, 1H), 4.90 (s, 1H), 3.76 -3.66 (m, 1H), 3.54-3.46 (m, peaks obscured by solvent), 3.28 - 3.22 (m, peaks obscured by solvent), 3.10 - 2.96 (m, 4H), 2.71 - 2.62 (m, 5H), 2.55 (dd, J = 1 1.8, 6.8 Hz, 1 H), 2.01 - 1.92 (m, 2H), 1.61 - 1.49 (m, 2H). LCMS-A: RT 1 .69 min; m/z 41 1.2 [M+H]\

Example 31 : 6-((1 -Acetylpiperidin-4-yl)amino)-W-(3-((2,3-dihydro-1 H-inden-2-yl)amino)- 2-h droxypropyl)pyrazine-2-carboxamide 52

(a) tert- Butyl ( 1 -acetylpiperidin-4-yl)carbamate A35

Acetic anhydride (4.71 mL, 49.9 mmol) was added to a solution of te f-butyl piperidin-4- ylcarbamate (10.0 g, 49.9 mmol) and triethylamine (10.4 mL, 74.9 mmol) in anhydrous DCM (100 mL) at 0 ' C. The reaction mixture was stirred at 0 °C for 2 hours before water (100 mL) and DCM (50 mL) were added. The organic phase was separated, washed with sat.

aqueous NaHCOa (100 mL) and dried (MgS0₄). The solvent was removed in vacuo to give the desired compound as a white solid (10.72 g, 89%). ¹H NMR (400 MHz, chloroform-cf) δ 4.54 - 4.42 (m, 2H), 3.80 - 3.70 (m, 1 H), 3.70 - 3.58 (m, 1 H), 3.12 (m. 1 H), 2.78 - 2.65 (m, 1 H), 2.07 (s, 3H), 2.05 - 1.97 (m, 1 H), 1.96 - 1.87 (m, 1 H), 1.43 (s, 9H), 1 .36 - 1 .20 (m, 2H).

(b) 1 -Acetylpiperidin-4-aminium chloride A36

A solution of iert-butyl (1-acetylpiperidin-4-y!)carbamate A35 (10.72 g, 44.24 mmol) in dioxane (100 mL) was cooled to 0 °C and treated with 4.0 M HCI in dioxane (12.2 mL, 48.7 mmol). The white precipitate was isolated by filtration and found to contain starting material. The precipitate was dissolved in MeOH (100 mL) and treated with 4.0 M HCI in dioxane (12.2 mL, 48.7 mmol) and the mixture was stirred at room temperature for 16 hours. Another aliquot of 4.0 M HCI in dioxane (6.10 mL, 24.4 mmol) was added and the reaction mixture was stirred for 1 .5 hours at 40 °C. The volatiles were removed in vacuo and the white solid was dried under high vacuum to give the desired compound (8.60 g, -90% purity, >95% yield). H NMR (400 MHz, DMSO-cfe) δ 8.52 - 8.23 (m, 3H), 4.39 - 4.26 (m, 1 H), 3.89 - 3.77 (m, 1 H), 3.28 - 3.14 (m, 1 H), 3.1 1 - 3.00 (m, 1 H), 2.65 - 2.54 (m, 1 H), 1.99 (s, 3H), 1.97 - 1.86 (m, 2H), 1.54 - 1.41 (m, 1 H), 1.41 - 1 .27 (m, 1 H).

(c) 6-Chloro-N-(3-((2,3-dihydro-1H-inden-2-yl)(2A-dimethoxybenz

hydroxypropyl)pyrazine-2-carboxamide A37

A mixture of 6-chloropyrazine-2-carboxylic acid (0.122 g, 0.771 mmol), DIPEA (0.24 mL, 1.4 mmol), HATU (0.293 g, 0.771 mmol) and DCM (10 mL) was stirred at room temperature under an atmosphere of nitrogen for 15 minutes. A solution of 1 -amino-3-((2.3-dihydro-1 H- inden-2-yl)(2,4-dimethoxybenzyl)amino)propan-2-ol 112 (0.250 g, 0.701 mmol) in DCM (5 mL) was then added and the mixture was stirred at room temperature overnight. The mixture was quenched by the addition of an aqueous NaOH solution (2 M, 20 mL) and H2O (10 mL). DCM (10 mL) was added and the mixture was separated. The aqueous was extracted with DCM (2 30 mL), the organic layers were combined and the solvent removed in vacuo. The resultant residue was purified by column chromatography (Biotage Isolera, 24 g S1O2 cartridge, 0-10% MeOH (containing 1 % v/v TEA) in DCM) to give the desired compound as a yellow oil (0.208 g, 60%). Ή NMR (400 MHz, chloroform-cf) δ 9.21 (s, 1 H), 8.74 (s, 1 H), 7.96 (s, 1 H), 7.20 - 7.13 (m, 2H), 7.10 - 7.02 (m, 3H), 6.49 - 6.32 (m, 2H), 4.31 - 4.09 (m, 1 H), 3.62 - 3.50 (m, 2H), 3.39 - 3.30 (m, 1 H), 3.12 - 2.89 (m, 4H), 2.80 (s, 6H), 2.57 - 2.38 (m, 2H), 1 .61 (s, 2H). LCMS-B: RT 3.66 min; m/z 497.3 [M+H]\

(d) 6-((1-Acetylpiperidin-4-yl)amino)-N-(3-((2,3-dihydro- 1H-inden-2-yl)(2.4- dimethoxybenzyl)amino)-2-hydroxypropyl)pyrazine-2-carboxamide A38

A mixture of 6-chloro-/V-(3-((2,3-dihydro-1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino)-2- hydroxypropyl)pyrazine-2-carboxamide A37 (0.108 g, 0.217 mmol), 1 -acetyipiperidin-4- aminium chloride A36 (0.058 g, 0.33 mmol), Cs₂C0₃ (0.354 g, 1.09 mmol), xantphos (0.006 g, 0.01 mmol) and Pd2(dba)a (0.010 g, 0.01 mmol) in 1.4-dioxane (5 mL) was bubbled with nitrogen for 10 min. The mixture was then stirred in the microwave at 120 C for 20 minutes. The mixture was diluted with EtOAc and the solids removed by filtration. The filtrate solvent was removed in vacuo and the residue was taken up in toluene (5 mL). An extra portion of 1 - acetylpiperidin-4-aminium chloride A36 (0.058 g, 0.33 mmol), rac-BINAP (0.014 g, 0.022 mmol), Pd₂(dba)₃ (0.020 g, 0.022 mmol), and i-BuOK (0.196 g, 1 .75 mmol) were added and the mixture was bubbled with nitrogen for 10 minutes before heating in the microwave at 120 C for 30 minutes. The solvent was removed in vacuo and the residue was purified by column chromatography (Biotage Isolera, 24 g S1O2 cartridge, 0-100% EtOAc in petroleum benzine 40-60 C then 0-20% MeOH in EtOAc) to give the desired compound as a yellow oil (0.032 g, 24%). LCMS-B: RT 3.32 min; m/z 603.4 [M+H]\

(e) 6-((1-Acetylpiperidin-4-yl)amino)-N-(3 (2,3-dihydro- 1H-inden-2-yl)amino)-2^

hydroxypropyl)pyrazine-2-carboxamide 52

A mixture of 6-((1 -acetylpiperidin-4-yl)amino)-/V-(3-((2,3-dihydro-1 H-inden-2-yl)(2,4- dimethoxybenzyl)amino)-2-hydroxypropyl)pyrazine-2-carboxamide A38 (0.032 g, 0.053 mmol) and TFA (5 mL) was stirred at 70 C for 16 hours. The mixture was cooled to room temperature and the volatiles were removed in vacuo. Water (30 mL) and aqueous NaOH (2 M) were added to the residue and the mixture was extracted with DCM (3 * 30 mL), dried (MgS04) and the solvent removed in vauco. The resultant oil was dissolved in a minimum amount of DCM and a solid was precipitated by the addition of cyclohexane. The solid was isolated by vacuum filtration and air dried to give the desired compound as a yellow solid (0.013 g, 54%, -90% purity as assessed by ¹H NMR). ¹H NMR (400 MHz. DMSO-cfe) δ 8.37 (t, J = 5.9 Hz, 1 H), 8.20 (s, 1 H), 8.09 (s, 1 H), 7.42 (d, J = 7.3 Hz, 1 H), 7.19 - 7.13 (m, 2H), 7.12 - 7.06 (m, 2H), 5.05 (s, 1 H), 4.25 - 4.16 (m. 1 H). 4.12 - 4.03 (m, 1 H), 3.85 - 3.71 (m, 4H), 3.28 - 3.16 (m, peaks obscured by solvent), 3.10 - 3.00 (m, 2H), 2.85 (t, J = 12.1 Hz, 1 H), 2.71 - 2.58 (m, 4H), 2.01 - 1 .98 (m, 3H), 1 .97 - 1.87 (m, 2H), 1.29 - 1.21 (m, peaks obscured by solvent). LCMS-B: RT 3.18 min: m/z 453.3 [M+H]\

Example 32: 5-((1 -AcetyIpipendin-4-yS)amino)-A/-(3-((2,3-dihydro-1 H-inden-2-yl)amino)- -hydroxypropyl)nicotinamide 53

(a) Methyl 5-bromonicotinate A39

A mixture of 5-bromonicotinic acid (1 .00 g, 4.95 mmol), sulfuric acid (98%, 0.5 mL) and MeOH (20 mL) was stirred at 80 °C for 16 hours. The volatiles were removed in vacuo and H2O (10 mL) and aqueous NaOH (2 M, 20 mL) were added to the residue. The aqueous was extracted with DCM (40 mL) and the mixture passed through an Isolute Phase Separator. The aqueous was extracted further with DCM (2 40 mL) with the mixture passed through an Isolute Phase Separator after each extraction. The organic layers were combined and the solvent removed in vacuo to give the desired compound as a yellow solid (0.822 g, 77%). ¹ H NM (400 MHz, chloroform-d) 6 9.12 (d, J = 1 .8 Hz, 1 H), 8.84 (d, J = 2.3 Hz, 1 H ), 8.45 - 8.42 (m, 1 H), 3.97 (s, 3H). LCMS-B: RT 3.56 min; m/z 216/218 [M+H]\

(b) Methyl 5-(( 1 -acetylpiperidin-4-yl)amino)nicotinate A40

A mixture of methyl 5-bromonicotinate A39 (0.500 g, 2.31 mmol), 1 -acetylpiperidin-4- aminium chloride A36 (0.620 g, 3.47 mmol), Cs₂C0₃ (3.77 g. 1 1 .6 mmol), xantphos (0.067 g, 0.12 mmol) and Pd₂(dba)₃ (0.106 g, 0.1 16 mmol) in 1 .4-dioxane (25 mL) was bubbled with nitrogen for 10 min. The mixture was stirred under an atmosphere of nitrogen at 80 C for 16 hours. Starting material was still present after this time, so another portion of CS2CO3 (3.77 g. 1 1 .6 mmol), xantphos (0.067 g, 0.12 mmol) and Pd₂(dba)₃ (0.106 g, 0.1 16 mmol) were added. The mixture was stirred under an atmosphere of nitrogen at 80 °C for 5 days. The mixture was returned to room temperature and diluted with EtOAc (50 mL), filtered and the filtrate was concentrated in vacuo. The crude solid was purified by column chromatography (Biotage Isolera, 40 g Si0₂ cartridge, 0-100% EtOAc in petroleum benzine 40-60 C, then 0- 20% MeOH in EtOAc) to give the desired compound as a yellow solid (0.273 g, 43%). ¹H NMR (400 MHz, DMSO-cfe) δ 8.26 (d, J = 1.8 Hz, 1 H), 8.20 (d, J = 2.8 Hz, 1 H), 7.41 - 7.39 (m, 1 H), 6.18 (d, J = 8.1 Hz, 1 H), 4.24 - 4.17 (m, 1 H), 3.84 (s, 3H), 3.81 - 3.73 (m, 1 H), 3.64 - 3.54 (m, 1 H), 3.25 - 3.15 (m, 1 H), 2.88 - 2.79 (m, 1 H), 2.01 (s, 3H), 1 .96 - 1 .83 (m, 2H), 1.36 - 1.19 (m, 2H). LCMS-B: RT 3.06 min; m/z 278.2 [M+H]\ (c) 5-(( 1 -Acetylpiperidin-4-yl)amino)nicotinic acid A41

A mixture of methyl 5-((1 -acetylpiperidin-4-yl)amino)nicotinate A40 (0.273 g, 0.984 mmol), LiOH.H₂0 (0.413 g, 9.84 mmol), THF (7 mL), MeOH (7 mL) and H₂0 (1 .5 mL) was stirred at 40 °C for 3 hours. The mixture was returned to room temperature and the volatiles were removed in vacuo. Water (20 mL) was added and the pH was adjusted to ~6 with aqueous HCI (2 M). A yellow precipitate was removed by filtration and the aqueous filtrate was passed through an Oasis HLB 35 cc LP extraction cartridge (6 g). The cartridge was washed with 3 column volumes of water before the lipophilic component was eluted with 3 column volumes of MeOH. Evaporation of the MeOH in vacuo gave the desired compound as a yellow oil (0.144 g, 56%). ¹H NMR (400 MHz, DMSO-de) δ 8.27 - 8.22 (m, 2H), 8.15 (d, J = 2.8 Hz, 1 H), 7.40 - 7.36 (m, 1 H), 6.07 (d, J = 8.0 Hz, 1 H), 4.26 - 4.16 (m, 1 H), 3.80 - 3.74 (m, 1 H), 3.60 - 3.53 (m, peaks obscured by solvent), 3.24 - 3.16 (m, peaks obscured by solvent), 2.87 - 2.77 (m, 1 H), 2.00 (s, 3H), 1.96 - 1.84 (m, 2H), 1 .36 - 1 .16 (m, 2H). LCMS- B: RT 1.80 min; m/z 264.2 [M+H]\ (d) 5-((1-Acetylpiperidin-4-yl)amino)-N-(3-((2,3-dihydro- 1H-inden-2-yl)(2.4- dimethoxybenzyl)amino)-2-hydroxypropyl)nicotinamide A42

A mixture of 5-((1 -acetylpiperidin-4-yl)amino)nicotinic acid A41 (0.050 g, 0.19 mmol), DIPEA (60 pL, 0.35 mmol) and HATU (0.072 g, 0.19 mmol) in DCM (5 mL) was stirred for 15 minutes at room temperature under an atmosphere of nitrogen. A solution of 1-amino-3- ((2,3-dihydro-1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino)propan-2-ol 112 (0.062 g, 0.17 mmol) in DCM (5 mL) was then added and the mixture was stirred for 4 hours at room temperature. Aqueous NaOH (2 M, 20 mL) was added and the aqueous was extracted with DCM (3 x 30 mL). The organic layers were combined, filtered through a Biotage phase separator and the volatiles removed in vacuo. The residue was purified by column chromatography (Biotage Isolera, 24 g Si0₂ cartridge, 0-20% MeOH in DCM) to give the desired compound as a colourless oil (0.066 g, 64%). LCMS-B: RT 3.51 min; m/z 602.4 [M+H]\

(e) 5-((1-Acetylpiperidin-4-yl)amino)-N-(3-((2,3-dihydro- 1H-inden-2-yl)amin

hydroxypropyl)nicotinamide 53

A mixture of 5-((1 -acetylpiperidin-4-yl)amino)-/V-(3-((2.3-dihydro-1 /+inden-2-yl)(2.4- dimethoxybenzyl)amino)-2-hydroxypropyl)nicotinamide A42 (0.066 g, 0.1 1 mmol) and TFA (5 mL) was stirred at 70 C for 4 hours. The mixture was cooled to room temperature and the volatiles were removed in vacuo. Water (30 mL) was added to the residue and the aqueous was washed with DCM (3 * 20 mL). The aqueous layer was made basic by the addition of aqueous NaOH (2 M) and the mixture was extracted with DCM (3 * 30 mL). The organic extract was filtered through a Biotage phase separator cartridge and the solvent was removed in vacuo. The colourless oil was dissolved in a minimum amount of DCM and a solid was precipitated by the addition of cyclohexane. The solid was isolated by vacuum filtration and air dried to give the desired compound as a white solid (0.040 g, 81 %). ¹H NMR (400 MHz, DMSO-cf_e) δ 8.50 (t, J = 5.7 Hz, 1 H), 8.17 (d, J = 1 .7 Hz, 1 H), 8.08 (d, J = 2.8 Hz, 1 H), 7.32 - 7.26 (m, 1 H), 7.19 - 7.14 (m, 2H), 7.12 - 7.07 (m, 2H), 6.02 (d, J = 8.0 Hz, 1 H), 4.96 - 4.84 (m, 1 H), 4.27 - 4.17 (m, 1 H), 3.82 - 3.75 (m, 1 H), 3.73 - 3.67 (m, 1 H), 3.60 - 3.53 (m, 1 H), 3.52 - 3.46 (m, 1 H), 3.29 - 3.14 (m, peaks obscured by solvent), 3.09 - 3.00 (m, 2H), 2.85 - 2.77 (m, 1 H), 2.69 - 2.61 (m, 3H), 2.57 - 2.52 (m, peaks obscured by solvent), 2.00 (s, 3H), 1.93 - 1.84 (m, 2H), 1.37 - 1.15 (m, 2H). LCMS-B: RT 3.10 min; m/z 452.3 [M+H]\

Example 33: 2-((1 -Acetylpiperidin4-yl)amino)-A -(3-((2,3-dihydro-1 H-inden-2-yl)amino)- 2-h droxypropyl)pyrimidine-4-carboxamide 54

(a) 2-Chloro-N-(3-((2,3-dihydro- 1H-inden-2-yl)(2,4-dimethoxybenzyl)amino)-2- hydroxypropyl)pyrimidine-4-carboxamide A43

A mixture of 2-chloropyrimidine-4-carboxylic acid (0.122 g, 0.771 mmol), DIPEA (0.24 mL, 1.4 mmol), HATU (0.293 g, 0.771 mmol) and DCM (10 mL) was stirred at room temperature under an atmosphere of nitrogen for 15 minutes. A solution of 1 -amino-3-((2.3-dihydro-1 H- inden-2-yl)(2.4-dimethoxybenzyl)amino)propan-2-ol 112 (0.250 g, 0.701 mmol) in DCM (5 mL) was then added and the mixture was stirred at room temperature overnight. The mixture was quenched by the addition of an aqueous NaOH solution (2 M, 20 mL) and H₂0 (10 mL). DCM (10 mL) was added and the mixture was separated. The aqueous was extracted with DCM (2 30 mL), the organic phases were combined and the solvent removed in vacuo. The resultant residue was purified by column chromatography (Biotage Isolera, 24 g S1O2 cartridge, 0-10% MeOH (containing 1 % v/v TEA) in DCM) to give the desired compound as a yellow oil (0.253 g, 73%). LCMS-B: RT 3.67 min; m/z 497.3 [M+H]\

(b) 2-(( 1 -Acetylpiperidin-4-yl)amino)-N-(3-((2, 3-di ydro- 1 H-inden-2-yl)(2,4- dimethoxybenzyl)amino)-2-hydroxypropyl)pyrimidine-4-carboxamide A44

A mixture of 2-chloro-/V-(3-((2, 3-dihydro- 1 /-/-inden-2-yl)(2,4-dimethoxybenzyl)amino)-2- hydroxypropyl)pyrimidine-4-carboxamide A43 (0.100 g, 0.201 mmol), 1-acetylpiperidin-4- aminium chloride A36 (0.108 g, 0.604 mmol), triethylamine (0.28 mL, 2.0 mmol) and /-PrOH (5 mL) was heated in the microwave at the following temperatures for the amount of time indicated, with a TLC of the reaction mixture conducted after each heating period: 1 10 C for 10 minutes; 1 10 °C for 15 minutes; 120 C for 15 minutes; 120 °C for 15 minutes; 130 C for 10 minutes; 150 C for 10 minutes. The solvent was removed in vacuo and the residue was purified by column chromatography (Biotage Isolera, 24 g S1O2 cartridge, 0-10% MeOH (containing 1 % v/v TEA) in DCM) to give the desired compound as a yellow oil (0.022 g, 18%). LCMS-B: RT 3.33 min; m/z 603.5 [M+H]\

(c) 2-((1-Acetylpiperidin-4-yi)amino)-N-(3-((2,3-dihydro- hydroxypropyl)pyrimidine-4-carboxamide 54

A solution of 2-((1-acetylpiperidin-4-yl)amino)-/V-(3-((2, 3-dihydro- 1 H-inden-2-yl)(2.4- dimethoxybenzyl)amino)-2-hydroxypropyl)pyrimidine-4-carboxamide A44 (0.022 g, 0.037 mmol) in TFA (5 mL) was stirred at 70 °C for 16 hours. The volatiles were removed in vacuo before the residue was diluted with H2O (10 mL). The aqueous phase was washed with DCM (3 x 30 mL) and then made basic by the addition of an aqueous NaOH solution (2 M). The aqueous was extracted with DCM (3 * 20 mL), the organic layers were combined, washed with brine, dried (MgSC ) and the solvent removed in vacuo. The crude residue was dissolved in a minimum amount of DCM and a precipitate formed by the addition of cyclohexane. The precipitate was isolated by vacuum filtration and air dried to give the desired compound as a white solid (0.006 g, 36%). ¹H NMR (400 MHz, DMSO-cfe) δ 8.60 - 8.24 (m, 2H), 7.46 (s, 1 H), 7.20 - 7.14 (m, 2H), 7.12 - 7.04 (m, 3H), 5.04 (s, 1 H), 4.27 (d, J = 13.3 Hz, 1 H), 4.04 (s, 1 H), 3.86 - 3.63 (m, 2H), 3.57 - 3.41 (m, peaks obscured by solvent), 3.19 - 2.99 (m. 3H), 2.80 - 2.57 (m, 6H), 2.00 (s, 3H), 1 .95 - 1 .81 (m, 2H), 1 .45 - 1.24 (m, peaks obscured by solvent). LCMS-A: RT 4.41 min; m/z 453.3 [M+H]⁺. Example 35: (fl)-2-((1 -Acetylpiperidin-4-yl)amino)-/V-{3-{(2,3-dihydro-1 H-inden-2- yl)amino)-2-hydroxypropyl)isonicotinamide 55

(a) Methyl 2-bromoisonicotinate A45

A solution of 2-bromoisonicotinic acid (5.00 g, 24.8 mmol) in MeOH (50 mL) was treated with sulfuric acid (98%, 0.50 mL, 9.4 mmol) and the reaction mixture was stirred at 80 "C for 1 hour. The mixture was returned to room temperature and stirred for a further 96 hours before heating to 80 °C and stirring for 24 hours. The reaction mixture was cooled to room temperature, and the volatiles were removed in vacuo. Aqueous NaOH (2 M, 50 mL) was added to the residue and the aqueous was extracted with EtOAc (3 * 50 mL). The organic layers were combined, washed with brine, dried (MgSC ) and the solvent removed in vacuo to give the desired compound as a yellow oil (4.14 g, 77%). ¹H NMR (400 MHz, chloroform- cf) δ 8.52 (dd, J = 5.0, 0.8 Hz, 1 H), 8.04 (t, J = 1 .2 Hz, 1 H), 7.80 (dd, J = 5.0, 1 .4 Hz, 1 H), 3.96 (s, 3H). LCMS-B: RT 3.55 min; m/z 216/218 [M+H]\

(b) Methyl 2-((1-acetylpiperidin-4-yl)amino)isonicotinate A46

A mixture of 1 -acetylpiperidin-4-aminium chloride A36 (2.25 g, 12.6 mmol), methyl 2- bromoisonicotinate A45 (1.81 g, 8.38 mmol), CS2CO3 (10.92 g, 33.51 mmol), xantphos (0.242 g, 0.419 mmol) and Pd₂(dba)₃ (0.384 g, 0.419 mmol) in 1 ,4-dioxane (40 mL) was bubbled with nitrogen for 10 min. The mixture was then stirred under an atmosphere of nitrogen at 80 C for 24 hours. Another portion of CS2CO3 (5.46 g, 16.8 mmol), xantphos (0.121 g, 0.209 mmol) and Pd₂(dba)₃ (0.192 g, 0.210 mmol) were added and the mixture was stirred under an atmosphere of nitrogen at 80 °C for 5 days. The reaction mixture was returned to room temperature and diluted with EtOAc (150 mL). Solid impurities were removed by filtration and the filtrate solvent was removed in vacuo. The resultant solid was purified by column chromatography (Biotage Isolera, 40 g S1O2 cartridge, 0-100% EtOAc in petroleum benzine 40-60 C, then 0-20% MeOH (containing 1 % v/v TEA) in EtOAc) to give the desired compound as a yellow solid (0.558 g, 24%). LCMS-B: RT 3.05 min; m/z 278.2 [M+H]\

(c) 2-(( 1 -A cetylpiperidin -4-yl)amino) i so nicotinic acid A47

A mixture of methyl 2-((1-acetylpiperidin-4-yl)amino)isonicotinate A46 (0.558 g, 2.01 mmol), UOH. H2O (1.69 g, 40.2 mmol), THF (7 mL), MeOH (7 mL) and H₂0 (1.5 mL) was stirred at 40 °C for 2 hours. The mixture was returned to room temperature and the volatiles were removed in vacuo. Water (30 mL) was added and the pH was adjusted to ~6 with aqueous HCI (2 M). The aqueous was passed through an Oasis HLB 35 cc LP extraction cartridge (6 g) which was washed with 4 column volumes of water. The lipophilic component was then eluted with 4 column volumes of MeOH. Evaporation of the MeOH in vacuo gave the desired compound as a yellow solid (0.197 g, 37%). The aqueous phase from the first iteration of cartridge purification was passed through another Oasis HLB 35 cc LP extraction cartridge (6 g). The column was washed with 4 column volumes of water and the product was eluted with 4 column volumes of MeOH. Evaporation of the MeOH in vacuo gave the desired compound as a white solid (0.128 g, 24%; overall yield: 0.325 g, 61 %). ¹H NMR (400 MHz, DMSO-ofe) δ 8.08 (d, J = 5.2 Hz, 1 H), 6.98 (s, 1 H), 6.86 - 6.79 (m, 2H), 4.26 - 4.16 (m, 1 H), 4.03 - 3.90 (m, 1 H), 3.81 - 3.72 (m, 1 H), 3.21 - 3.12 (m, 1 H), 2.85 - 2.74 (m, 1 H), 2.00 (s, 3H), 1 .97 - 1 .83 (m, 2H), 1 .41 - 1 .16 (m, 2H); -OH not observed. LCMS-B: RT 1.17 min; m/z 262.1 [M-H]-.

(d) (R)-2-((1-Acetylpiperidin-4-yl)amino)-N-(3-((2,3-dihydro- 1H-inden-2-yl)ami

hydroxypropyl)isonicotinamide 55

A mixture of 2-((1 -acetylpiperidin-4-yl)amino)isonicotinic acid A47 (0.067 g, 0.26 mmol), DIPEA (63 μΙ_, 0.36 mmol), HATU (0.097 g, 0.26 mmol) and DCM (5 mL) was stirred at room temperature for 15 minutes. A solution of (S)-1-amino-3-((2.3-dihydro-1 H-inden-2- yl)amino)propan-2-ol I4 (0.050 g, 0.24 mmol) in DCM (5 mL) was then added and the mixture was stirred for 16 hours at room temperature. Water (20 mL) was added and the aqueous was extracted with DCM (3 * 30 mL). The organic layers were combined, washed with brine, dried (MgS0₄) and the solvent removed in vacuo. The resultant residue was purified by column chromatography (Biotage Isolera, 24 g S1O2 cartridge, 0-30% MeOH (containing 1 % v/v TEA) in DCM) to give a colourless oil. The oil was taken up in a minimum amount of DCM and the product was precipitated by the addition of cyclohexane. The product was isolated by vacuum filtration and air dried to give the desired compound as a white solid (0.01 1 g, 10%). ¹ H NMR (400 MHz, DMSO-c ₆) δ 8.49 (t, J = 5.7 Hz, 1 H), 8.02 (d, J = 5.2 Hz, 1 H), 7.21 - 7.14 (m, 2H), 7.12 - 7.06 (m, 2H), 6.84 (s, 1 H), 6.77 (d, J = 5.5 Hz, 1 H), 6.72 (d, J = 7.6 Hz, 1 H), 4.93 (s, 1 H), 4.28 - 4.14 (m, 1 H), 4.02 - 3.88 (m, 1 H), 3.83 - 3.66 (m, 2H), 3.52 (p, J = 6.6, 6.6, 6.5, 6.5 Hz, 1 H). 3.29 - 3.12 (m. 3H), 3.1 1 - 3.00 (m, 2H), 2.82 - 2.74 (m, 1 H), 2.72 - 2.62 (m, 3H). 2.59 - 2.53 (m, 1 H), 2.00 (s, 3H), 1 .96 - 1 .81 (m, 2H), 1 .38 - 1 .14 (m, 2H); N-H not observed. LCMS-B: RT 3.1 1 min; m/z 452.3 [M+H]\ Example 36: (S)-2-((1 -Acetyipiperidin- -yl)amino)-N-(3-((2,3-dihydro-1 H-inden-2- l)amino)-2-hydroxypropyl)isonicotinamide 56

(a) (R)-2-((1-Acetylpiperidin-4-yl)amino)-N-(3-((2,3-dihydro- 1H-inden-2-yl)(2

dimethoxybenzyl)amino)-2-hydroxypropyl)isonicotinamide A48

A mixture of 2-((1 -acetylpiperidin-4-yl)amino)isonicotinic acid A47 (0.075 g, 0.29 mmol), DIPEA (68 pL, 0.39 mmol), HATU (0.108 g, 0.285 mmol) and DCM (10 mL) was stirred at room temperature for 15 minutes. A solution of (f?)-1 -amino-3-((2,3-dihydro-1 H-inden-2- yl)(2,4-dimethoxybenzyl)amino)propan-2-ol 110 (0.092 g, 0.26 mmol) in DCM (5 mL) was then added and the mixture was stirred for 24 hours at room temperature. Water (30 mL) was added and the aqueous was extracted with DCM (3 * 30 mL). The organic layers were combined, washed with brine, dried (MgS0₄) and the solvent removed in vacuo. The resultant residue was purified by column chromatography (Biotage Isolera, 12 g S1O2 cartridge, 0-20% MeOH (containing 1 % v/v TEA) in DCM) to give the desired compound as a colourless oil (0.084 g, 54%). LCMS-B: RT 3.28 min; m/z 602.5 [M+H]\ (b) (S)-2-((1 -Acetylpiperidin-4-yl)amino)^-(3-((2,3-dihydro-1H-inden-2-yl)amino)-2- hydroxypropyl)isonicotinamide 56

A mixture of (/?)-2-((1-acetylpiperidin-4-yl)amino)-/V-(3-((2,3-dihydro-1 H-inden-2-yl)(2,4- dimethoxybenzyl)amino)-2-hydroxypropyl)isonicotinamide A48 (0.080 g, 0.13 mmol) and TFA (5 mL) was stirred at 70 C for 16 hours. The volatiles were removed in vacuo before the residue was diluted with H₂0 (20 mL). The aqueous phase was washed with DCM (3 * 30 mL) and then made basic by the addition of solid NaHCC<3. The aqueous was extracted with DCM (3 x 30 mL), the organic layers were combined, washed with brine, dried (MgSCu) and the solvent removed in vacuo. The resultant residue was purified by column

chromatography (Biotage Isolera, 24 g S1O2 cartridge, 0-40% MeOH (containing 1 % v/v TEA) in DCM) to give a colourless oil. The oil was dissolved in a minimum amount of DCM and the product was precipitated by the addition of cyclohexane. The solid was isolated by vacuum filtration and air dried to give the desired compound as a white solid (0.030 g, 50%). ¹H NMR (400 MHz, DMSO-cfe) δ 8.48 (t, J = 5.7 Hz, 1 H), 8.02 (d, J = 5.3 Hz, 1 H), 7.21 - 7.14 (m, 2H), 7.13 - 7.06 (m, 2H), 6.84 (s, 1 H), 6.77 (d, J = 5.3 Hz, 1 H), 6.72 (d, J = 7.6 Hz, 1 H), 4.92 (s, 1 H), 4.27 - 4.15 (m, 1 H), 4.04 - 3.89 (m, 1 H), 3.83 - 3.66 (m, 2H), 3.58 - 3.47 (m, 1 H), 3.30 - 3.22 (m, peaks obscured by solvent), 3.20 - 3.12 (m, 1 H), 3.10 - 3.00 (m, 2H), 2.84 - 2.74 (m, 1 H), 2.72 - 2.62 (m, 3H), 2.60 - 2.52 (m, 1 H), 2.00 (s, 3H), 1.97 - 1.82 (m, 2H), 1.38 - 1 .15 (m, 2H). LCMS-B: RT 3.09 min; m/z 452.3 [M+H]\

Examaple 37: 6-((1 -Acetylpiperidin-4-yl)amino)-/V-(3-((2,3-dihydro-1 H-inden-2- yl amino)-2-hydroxypropyl)picolinamide 57

(a) Ethyl 6-(( 1 -acetylpiperidin-4-yl)amino)picolinate A49

A mixture of 1 -acetylpiperidin-4-aminium chloride A36 (0.231 g, 1 .29 mmol), 6- bromopyridine-2-carboxylic acid ethyl ester (0.198 g, 0.862 mmol), CS2CO3 (1 .40 g, 4.31 mmol), xantphos (0.025 g, 0.043 mmol) and Pd₂(dba)₃ (0.039 g, 0.043 mmol) in 1 ,4-dioxane (10 mL) was bubbled with nitrogen for 10 min. The mixture was then stirred under an atmosphere of nitrogen at 80 °C for 16 hours. The reaction mixture was returned to room temperature and diluted with MeOH (20 mL). Solid impurities were removed by filtration and the filtrate solvent was removed in vacuo. The resultant solid was purified by column chromatography (Biotage I solera, 24 g S1O2 cartridge, 0-100% EtOAc in petroleum benzine 40-60 °C, then 0-10% MeOH in EtOAc) to give the desired compound as a yellow oil (0.121 g, 48%). LCMS-A: RT 4.65 min; m/z 292.2 [M+H]\

(b) 6-(( 1 -Acetylpiperidin-4-yl)amino)picolinic acid A50

A mixture of ethyl 6-((1-acetylpiperidin-4-yl)amino)picolinate A49 (0.121 g, 0.415 mmol), UOH.H2O (0.349 g, 8.31 mmol), MeOH (7 mL), THF (7 mL) and H₂0 (1.5 mL) was stirred at room temperature for 16 hours. The volatiles were removed in vacuo and the resultant solid was taken up in H2O (10 mL) and the aqueous adjusted to pH ~6 with aqueous HCI (2 M). The mixture was loaded onto an Oasis HLB 35 cc LP extraction cartridge (6 g) which was washed with 3 column volumes of water. The lipophilic component was then eluted with 3 column volumes of MeOH. Evaporation of the MeOH in vacuo gave the desired compound as a yellow solid (0.097 g, 89%). H NMR (400 MHz. chloroform-cf) δ 7.91 - 7.82 (m, 1 H), 7.46 (d, J = 7.2 Hz, 1 H), 6.93 (d, J = 8.4 Hz, 1 H), 4.25 (s, 1 H), 3.99 - 3.88 (m, 1 H), 3.80 (s, 1 H), 3.41 - 3.31 (m, 1 H), 3.29 - 3.14 (m, 1 H), 2.13 (s, 3H), 2.09 - 1.94 (m, 2H), 1.83 (s, 1 H), 1.74 - 1.60 (m, 1 H). LCMS-A: RT 1 .75 min; m/z 264.2 [M+H]\

(c) 6-((1-Acetylpiperidin-4-yl)amino)-N-(3-((2,3-dihy

hydroxypropyl)picolinamide 57

A mixture of 6-((1 -acetylpiperidin-4-yl)amino)picolinic acid A50 (0.097 g, 0.37 mmol), triethylamine (0.20 mL, 1 .5 mmol), and DCM (5 mL) was cooled to 0 C under a nitrogen atmosphere before isobutyl chloroformate (48 L, 0.37 mmol) was added. The mixture was stirred for 20 min at 0 C before being transferred via syringe to a solution of 1-amino-3- ((2.3-dihydro-1 H-inden-2-yl)amino)propan-2-ol I2 (0.076 g, 0.37 mmol) in DCM (5 mL) at 0 C. After 2 h the mixture was diluted with DCM (10 mL), water (10 mL) and aqueous sodium hydroxide (2 M, 10 mL). The separated aqueous phase was extracted with DCM (2 15 mL), the pooled DCM extracts washed with brine (25 mL), dried (MgS0₄) and the solvent evaporated in vacuo. The residue was purified by column chromatography (Biotage Isolera, 12 g Si0₂ cartridge, 0-20% MeOH (containing 1 % v/v 2 M NH₃ in MeOH) in DCM) to give the desired compound as a white solid (0.049 g. 29%). Ή NMR (400 MHz, DMSO-cfe) δ 8.66 - 8.55 (m, 1 H), 7.57 - 7.47 (m, 1 H), 7.28 - 7.21 (m, 2H), 7.20 - 7.14 (m, 3H), 6.82 (d, J = 7.7 Hz, 1 H), 6.67 (d, J = 8.3 Hz, 1 H), 5.92 - 5.81 (m, 1 H), 4.27 - 4.14 (m, 2H). 4.09 - 3.96 (m, 2H), 3.82 - 3.67 (m, 1 H), 3.52 - 3.38 (m, 2H), 3.30 - 3.21 (m, 3H), 3.18 - 3.03 (m, 3H). 2.96 - 2.84 (m, 2H), 2.03 - 1 .96 (m, 3H), 1 .96 - 1 .83 (m, 2H), 1 .38 - 1.20 (m, 2H); N-H not observed. LCMS-A: RT 4.46 min; m/z 452.3 [M+H]\

Example 38: 6-((1 -Acetylazetidin-3-yl)amino)-A -{3-((2,3-dihydro-1 H-inden-2-yl)amino)- -hydroxypropyl)pyrimidine-4-carboxamide 2,2,2-trifluoroacetic acid salt 58

(a) tert-Butyl ( 1 -acetylazetidin-3-yl)carbamate A51

Acetic anhydride (0.81 mL, 8.6 mmoi) was added to a solution of iert-butyl azetidin-3- ylcarbamate (0.990 g, 5.75 mmol) and TEA (1.20 mL, 8.62 mmol) in DCM (20 mL) at 0 °C. The mixture was stirred for 2 hours at 0 ' C before water (50 mL) was added. The organic phase was separated and the aqueous was extracted with DCM (2 25 mL). The organic layers were combined, washed with brine and dried (MgSC ). The solvent was removed in vacuo to give the desired compound as a colourless oil (0.915 g, 74%). ¹H NMR (400 MHz, chloroform-d) 5 5.13 - 4.92 (m, 1 H), 4.55 - 4.34 (m, 2H), 4.28 (t, J = 9.2 Hz, 1 H), 3.94 (dd, J = 8.7, 4.8 Hz, 1 H), 3.79 (dd, J = 10.4, 4.9 Hz, 1 H), 1 .86 (s, 3H), 1.44 (s, 9H).

(b) 1 -(3-Aminoazetidin- 1 -yl)ethan- 1 -one A52

A solution of iert-butyl (1-acetylazetidin-3-yl)carbamate A51 (0.879 g, 4.10 mmol) in MeOH (20 mL) was stirred with HCI (-1.25 M in MeOH, 3.61 mL, 4.51 mmol) at room temperature for 30 minutes. Another aliquot of HCI (-1 .25 M in MeOH, 3.61 mL, 4.51 mmol) was added and the mixture was stirred for a further 16 hours at room temperature. TLC showed the continued presence of starting material, so another aliquot of HCI (-1.25 M in MeOH, 7.22 mL, 9.03 mmol) was added. The mixture was stirred for 2 hours at room temperature and then at 45 °C for 2 hours. The solvent was evaporated under reduced pressure and the resultant residue was taken up in DCM (10 mL) and treated with TFA (3.14 mL, 41.0 mmol). The mixture was stirred for 3 hours at room temperature before the voiatiles were removed in vacuo. The residue was taken up in MeOH and loaded onto an SCX cartridge (BondElut, 10 g). The column was washed with 5 column volumes of MeOH before the desired product was eluted with 5 column volumes of 3.5 M NH3 in MeOH. The basic fractions were combined and the solvent removed in vacuo to give the desired compound as a yellow oil (0.505 g, -90% purity, >95% yield). H NMR (400 MHz, chloroform-d) δ 4.39 - 4.29 (m, 1 H), 4.27 - 4.19 (m, 1 H), 3.90 - 3.72 (m, 2H), 3.70 - 3.59 (m, 1 H), 1 .86 (s, 3H); -NH₂ not observed.

(c) 6-Chloropyrimidine-4-carbonyl chloride A53

Oxalyl chloride (15.3 mL, 0.178 mol) was added to a mixture of 6-hydroxypyrimidine-4- carboxylic acid (5.00 g, 35.7 mmol) in EtOAc (50 mL). The mixture was stirred at room temperature for 20 min before DMF (0.1 mL) was added and stirring continued at 80 C for 16 hours. The volatiles were removed in vacuo and the residue was azeotroped with toluene (x 3) to give the desired compound as a black solid (5.87 g, 93%). ¹ H NMR (400 MHz, DMSO-cf₆) δ 9.23 (d, J = 1.1 Hz, 1 H), 8.15 (d, J = 1 .1 Hz, 1 H).

(d) 6-Chloro-N-(3-((2,3-dihydro- 1H-inden-2-yl)amino)-2-hydroxypropyl)pyhmidine-4- carboxamide A54

A solution of 6-chloropyrimidine-4-carbonyl chloride A53 (0.179 g, 1 .01 mmol) in DCM (10 mL) was slowly added to a mixture of 1 -amino-3-((2.3-dihydro-1 /-/-inden-2-yl)amino)propan- 2-ol I2 (0.250 g, 1 .21 mmol) and TEA (1 .41 mL. 10.1 mmol) in DCM (10 mL) at 0 C under an atmosphere of nitrogen. The mixture was slowly allowed to return to room temperature and stirred for a further 16 hours under a balloon of nitrogen. Water (20 mL) and an aqueous NaOH solution (2 M, 10 mL) were added and the mixture was extracted with DCM (3 * 30 mL). The organic layers were combined, washed with brine and dried (MgSO.i). The solvent was removed in vacuo and the residue was purified by column chromatography (Biotage

Isolera, 24 g Si0₂ cartridge, 0-10% MeOH (containing 1 % v/v 2 M NH₃ in MeOH) in DCM) to give the desired compound as a white solid (0.1 1 1 g, 32%). Ή NMR (400 MHz, DMSO-cfe) δ 9.17 (d, J = 0.6 Hz, 1 H), 9.07 (t. J = 5.8 Hz, 1 H), 8.08 (d. J = 1.1 Hz, 1 H), 7.20 - 7.14 (m, 2H), 7.12 - 7.06 (m, 2H), 5.06 - 4.92 (m, 1 H), 3.80 - 3.68 (m, 1 H), 3.53 - 3.46 (m, 1 H), 3.45 - 3.38 (m, peaks obscured by solvent), 3.10 - 3.00 (m, 2H), 2.70 - 2.57 (m, 4H). LCMS-A: RT 4.46 min; m/z 347.1 [M+H]⁺.

(e) 6-((1-Acetylazetidin-3-yl)amino)^-(3-((2.3-dihydro- 1H-inden-2-yl)amino)-2- hydroxypropyl)pyrimidine-4-carboxamide 2, 2, 2-thfluoroacetic acid salt 58

A mixture of 1 -(3-aminoazetidin-1-yl)ethan-1-one A52 (0.013 g, -90% purity, 0.10 mmol), 6- chloro-A/-(3-((2,3-dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)pyrimidine-4-carboxamide A54 (0.035 g, 0.10 mmol), triethylamine (0.056 ml_, 0.40 mmol) and /-PrOH (5 mL) was stirred at 70 C for 1 hour and then 75 C for 60 hours. The mixture was returned to room temperature and the solvent was evaporated under reduced pressure. The residue was purified by column chromatography (Biotage Isolera, 4 g S1O2 cartridge, 0-20% MeOH (containing 1 % v/v NEt.3) in DCM). The resultant white solid was purified further by RP-HPLC (5-100% CH3CN in water containing 0.1 % TFA over 20 minutes) to give the TFA salt of the desired compound (0.020 g, 37% yield). ¹H NMR (400 MHz, CD₃OD) δ 8.60 - 8.50 (m, 1 H),

7.29 - 7.24 (m, 2H). 7.23 - 7.17 (m, 3H), 4.83 - 4.72 (m, 1 H), 4.64 - 4.54 (m, 1 H), 4.40 -

4.30 (m, 1 H), 4.16 - 4.05 (m, 3H), 3.90 (dd, J = 10.4, 5.2 Hz, 1 H), 3.60 - 3.47 (m, 2H), 3.46 - 3.37 (m, 2H), 3.25 (dd, J = 12.6. 3.1 Hz, 1 H), 3.18 - 3.09 (m, 2H), 3.03 (dd, J = 12.8, 9.4

Hz, 1 H), 1.90 (s, 3H). LCMS-A: RT 1.67 min; m/z 459.1 [M+CI]\ 423.2 [M-H]\

Example 39: 6-((1 -Acetylpiperidin^-yl)amino)-A/^3^(2,3-dihydro-1 H-inden-2-yl)amino)^ 2-hydroxypropyl)pyrimidine-4-carboxamide 2,2,2-trifluoroacetic acid salt 59

A mixture of 1 -acetylpiperidin-4-aminium chloride A36 (0.026 g, 0.14 mmol), 6-chloro- V-(3- ((2,3-dihydro-1 /-/-inden-2-yl)amino)-2-hydroxypropyl)pyrimidine-4-carboxamide A54 (0.050 g, 0.14 mmol). triethylamine (0.080 mL. 0.58 mmol) and /-PrOH (5 mL) was stirred at 60 C for 16 hours. Another aliquot of triethylamine (0.16 mL, 1 .1 mmol) was added and the mixture was stirred at 60 °C for 3 hours and then at 70 C for 16 hours. Another aliquot of triethylamine (0.24 mL, 1 .7 mmol) was added and the mixture was stirred at 70 °C for 4.5 hours. A final addition of triethylamine (1.0 mL, 7.2 mmol) was followed by stirring of the reaction mixture for 16 hours at 75 °C. The solvent was evaporated under reduced pressure and the residue purified by column chromatography (Biotage Isolera, 4 g S1O2 cartridge, 0- 20% MeOH (containing 1 % v/v 2 M NH₃/MeOH) in DCM). The resultant white solid was purified further by RP-HPLC (5-100% CH₃CN in water containing 0.1 % TFA over 20 minutes) to give the TFA salt of the desired compound (0.026 g, 32% yield). ¹H NMR (400 MHz, CD3OD) δ 8.55 (s, 1 H), 7.28 - 7.24 (m, 2H), 7.23 - 7.19 (m, 2H), 7.16 (s, 1 H), 4.47 (d, J = 13.4 Hz, 1 H), 4.34 - 4.18 (m, 1 H), 4.16 - 4.06 (m, 2H), 4.01 - 3.91 (m, 1 H), 3.59 - 3.47 (m, 2H), 3.46 - 3.37 (m, 2H), 3.28 - 3.22 (m, peak obscured by solvent), 3.20 - 3.09 (m, 2H), 3.09 - 2.98 (m, 1 H), 2.93 - 2.82 (m, 1 H), 2.13 (s, 3H), 2.12 - 1.98 (m, 2H), 1.61 - 1.39 (m, 2H); pyrimidine N-H, amide H-H, -OH and amine N-H not observed. LCMS-A: RT 4.28 min; m/z 487.2 [M+CI]\ 451.2 [M-H]\

Example 40: Amide couplings:

General Method X2

To a solution of the amine (0.20 mmol, 1 equiv) in DMF (2 mL) and MeCN (5 mL) was added 114 (100 mg, 0.198 mmol, 1 equiv), DIPEA (104 pL, 0.595 mmol, 3 equiv) and HATU (1 13 mg, 0.297 mmol, 1.5 equiv). The reaction was stirred at room temperature for 16 h. The mixture was quenched with a 1 M aqueous solution of sodium hydroxide (10 mL) and stirred for 2 h at room temperature. Dichloromethane was added and the mixture stirred and then passed through a phase separation cartridge (3 repeats). The organic filtrates were dried in vacuo. The residue was taken up in TFA (0.5 mL) and stirred at 70 C for 5 h. Water (2 mL) was added and the aqueous was washed with DCM (3 * 2 mL) and the layers separated using a phase separation cartridge. The aqueous was brought to pH ~ 14 by the addition of aqueous NaOH (1 M) and then extracted with DCM (2 2 mL) using a phase separation cartridge to isolate the organic fraction. The organic layers were concentrated in vacuo. The residue was taken up in MeOH and purified by solid-phase extraction (1 g SCX-2 cartridge, 3 column volumes of methanol followed by 3 column volumes of 0.2 M methanolic ammonia) to give the desired compound. eneral Method X3

To a solution of the acid (53 mg, 0.22 mmol, 1 equiv), in MeCN (3 ml_) and DMF (1 mL) was added DIPEA (1 17 μΙ_, 0.670 mmol, 3 equiv) and HATU (128 mg, 0.337 mmol, 1.5 equiv). A solution of the amine 110 (80 mg, 0.22 mmol, 1 equiv) in DMF (1 mL) was added and the solution stirred at room temperature for 16 h. The reaction was quenched with a 1 M aqueous solution of NaOH (15 mL) and then extracted with dichloromethane (3 ^χ 8 mL - separation using a phase separation cartridge). The pooled organic filtrates were concentrated in vacuo then TFA (0.5 mL) was added and the solution stirred at 70 C for 5 h. The reaction was quenched by the addition of DCM (2 mL) and water (2 mL). The phases were thoroughly mixed and the aqueous layer isolated using a phase separation cartridge. The aqueous layer was brought to pH ~ 14 by the addition of aqueous NaOH (1 M) and then extracted with DCM (2 2 mL) using a phase separation cartridge to isolate the organic fraction. The organic layers were concentrated in vacuo. The residue was taken up in MeOH and purified by solid-phase extraction (1 g SCX-2 cartridge, 3 column volumes of methanol followed by 3 column volumes of 0.2 M ammonia in methanol) to give the desired compound.

Example Name and Structure LCMS data Method

60 LCMS-B: RT = X2

3.13 min, m/z =

438 [M+H] '

(S)-/V-(3-((2,3-dihydro-1 H-inden-2- yl )amino)-2-hyd roxy propyl )-4-(4- hydroxypiperidine-1- carbonyl)benzamide

61 LCMS-B: RT = X2

3.14 min, m/z =

452 [M+H]'

(S)-A/-(3-((2,3-dihydro-1 /+inden-2- yl)amino)-2-hydroxypropyl)-4-(4- (hydroxymethyl)piperidine-l - carbonyl)benzamide

62 LCMS-A: RT = X3

3.39 min, m/z =

424

(S)-A/-(3-((2,3-dihydro-1 H-inden-2- yl)amino)-2-hydroxypropyl)-3- (morpholine-4-carbonyl)benzamide

Example 41 : A -(3-{(2,3-Dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)-4-(2-

(a) N-(3-((2,3-dihydro-1H-inden-2-yl)amino)-2-hydroxypropyl)-4-(2-oxopiperidin- 1 - yl)benzamide 63

1-Amino-3-((2.3-dihydro-1 H-inden-2-yl)amino)propan-2-ol I2 (0.084 g, 0.407 mmol), 4-(2- oxopiperidin-1 -yl)benzoic acid (0.074 g, 0.339 mmol). EDCI (0.098 g, 0.509 mmol), HOBt (0.069 g, 0.509 mmol) and DIPEA (0.177 mL, 1 .018 mmol) in dry DMF (5 mL) were stirred at room temperature for 45 hours under an atmosphere of nitrogen. The reaction was diluted with EtOAc (50 mL) and washed with a sat. aqueous NaHCC>3 (50 mL), the aqueous layer was extracted with EtOAc (50 mL) and the combined organic layers were washed with brine (50 mL), dried ( a2S0₄), filtered and concentrated in vacuo to give a beige solid. Purification by column chromatography (Isolera Biotage, S1O2 cartridge, 0-100% EtOAc in petroleum benzine 40-60 C, then 0-35% MeOH in EtOAc) gave a gum which was triturated with diethyl ether and further air-dried to yield the desired compound (0.010 g, 7% yield) as a white solid. H NMR (400 MHz, d₆-DMSO) δ 8.47 (t, J = 5.7 Hz, 1 H), 7.85 - 7.79 (m, 2H), 7.40 - 7.33 (m, 2H), 7.21 - 7.14 (m, 2H), 7.14 - 7.06 (m, 2H), 4.91 - 4.85 (m, 1 H), 3.76 - 3.66 (m, 1 H), 3.66 - 3.61 (m. 2H), 3.49 (p, J = 6.6 Hz, 1 H), 3.38 - 3.23 (m, 3H, obscured by solvent signal), 3.09 - 3.00 (m, 2H), 2.70 - 2.61 (m, 3H), 2.58 - 2.52 (m, 1 H), 2.44 - 2.38 (m, 2H), 1.93 - 1.80 (m, 4H). LCMS-B: RT 3.227 min, m/z 408.3 [M+H]⁺

Example 42: 4-((1 -Acetylpiperidin-4-yl)amino)-N-{3-{(2,3-dihydro-1 H-inden-2-yl) 2-h roxypropyl) benzamide 64

(a) tert-Butyl 4-((4-(methoxycarbonyl)phenyl)amino)piperidine- 1 -carboxylate A55

Methyl 4-aminobenzoate (0.250 g, 1.654 mmol) and ie t-butyl 4-oxopiperidine-1 -carboxylate (0.494 g, 2.481 mmol) were dissolved in dry DCE (15 mL) under an atmosphere of nitrogen and acetic acid (0.189 mL, 3.308 mmol) was added followed by sodium

triacetoxyborohydride (0.701 g, 3.308 mmol). The reaction was stirred at room temperature for 45 hours, then quenched by addition of water (5 mL) and diluted with EtOAc (50 mL) and sat. aqueous NaHCC>3 (50 mL). The organic phase was washed with brine (50 mL), dried (Na2S0₄), filtered and concentrated in vacuo then purified by silica gel column

chromatography (Isolera Biotage: 40 g S1O2 cartridge, 0-30% EtOAc in petroleum benzine 40-60 C) to give the desired compound (0.478 g, 86% yield) as a colourless oil which partly solidified upon standing. H NMR (400 MHz, CDCI₃) δ 7.92 - 7.84 (m, 2H), 6.74 - 6.53 (m, 2H), 4.16 - 3.98 (m, 2H), 3.86 (s, 3H), 3.48 (tt, J = 10.4, 3.9 Hz, 1 H), 2.95 - 2.82 (m, 2H), 2.08 - 1.99 (m, 2H), 1.49 - 1 .35 (m, 1 1 H). NH proton not visible. LCMS-B: RT 3.826 min; m/z 279.2 [M- <Bu + H]\ (b) Methyl 4-(piperidin-4-ylamino)benzoate A 56

tert-Butyl 4-((4-(methoxycarbonyl)phenyl)amino)piperidine-1-carboxylate A55 (0.475 g, 1 .420 mmol) was dissolved in DCM (10 mL) under an atmosphere of nitrogen and trifluoroacetic acid (3.2 mL, 42.6 mmol) was added. The reaction was then stirred at room temperature for 2 hours. The volatiles were removed in vacuo and the resulting residue diluted with EtOAc (50 mL) followed by sat. aqueous Na2CC>3 (50 mL). The layers were separated and the aqueous layer was extracted with EtOAc (50 mL), the combined organic layers were washed with water (50 mL), brine (50 mL), dried (Na2S0₄), filtered and concentrated in vacuo to give the desired compound (0.178 g, 53% yield) as a white solid. ¹H NMR (400 MHz, d₆-DMSO) δ 7.70 - 7.64 (m, 2H), 6.62 - 6.57 (m, 2H), 6.44 (d, J = 7.8 Hz, 1 H), 3.73 (s, 3H), 2.99 (dt, J = 12.8, 3.6 Hz, 2H), 2.60 (td, J = 12.1 , 2.6 Hz, 2H), 1.92 - 1.79 (m, 2H), 1 .35 - 1.21 (m, 2H). One of NH protons not visible, CH proton obscured by solvent peak. LCMS-B: RT 3.179 min; m/z 235.2 [M+H]\

(c) Methyl 4-((1 -acetylpiperidin-4-yl)amino)benzoate A57

Methyl 4-(piperidin-4-ylamino)benzoate A56 (0.175 g, 0.747 mmol) was dissolved in DCM (7 mL) under an atmosphere of nitrogen and DIPEA (0.260 mL, 1.494 mmol) was added followed by acetic anhydride (0.070 mL, 0.747 mmol). The reaction was stirred at room temperature for 6 hours, then diluted with EtOAc (50 mL) followed by sat. aqueous a2C03 (50 mL). The layers were separated and the aqueous layer was extracted with EtOAc (50 mL), the combined organic layers were washed with brine (50 mL), dried ( a2S0₄), filtered and concentrated in vacuo to give the desired compound (0.170 g, 82% yield) as an off- white solid.¹H NMR (400 MHz, d₆-DMSO) δ 7.71 - 7.66 (m, 2H), 6.67 - 6.60 (m, 2H), 6.46 (d, J = 7.8 Hz, 1 H), 4.28 - 4.20 (m, 1 H), 3.82 - 3.71 (m, 4H), 3.63 - 3.51 (m, 1 H), 3.22 - 3.12 (m, 1 H), 2.83 - 2.74 (m, 1 H), 2.00 (s, 3H), 1 .97 - 1 .84 (m, 2H), 1 .37 - 1.15 (m, 2H). LCMS-B: RT 3.333 and 3.376 (two major peaks present) min; m/z 277.2 [M+H]\

(d) 4-((1 -Acetylpiperidin-4-yl)amino)benzoic acid A58

Methyl 4-((1-acetylpiperidin-4-yl)amino)benzoate A57 (0.168 g, 0.608 mmol) was dissolved in THF (7 mL), MeOH (1 mL) and water (1 mL) and lithium hydroxide monohydrate (0.077 g, 1.823 mmol) was added. The reaction was then stirred at room temperature for 70 hours. Additional lithium hydroxide monohydrate (0.154 g) was added and the reaction was stirred for another 8 hours and then heated to 40 C and stirred for an additional 45 hours. The volatiles were removed in vacuo and the resulting residue diluted with EtOAc (50 mL) followed by 2M aqueous NaOH (50 mL). The layers were separated and the organic layer was washed with 2M aqueous NaOH (50 mL), the aqueous layers were combined and acidified by addition of 2 M aqueous HCI (checked with pH paper), then extracted with EtOAc (2 x 70 mL). The combined organic layers were washed with brine (100 mL), dried (Na2SO.i), filtered and concentrated in vacuo to give the desired compound (0.060 g, 38% yield) as an off-white solid. ¹ H NMR (400 MHz, d₆-DMSO) δ 12.04 (br s, 1 H), 7.69 - 7.62 (m, 2H), 6.65 - 6.57 (m, 2H), 6.36 (d, J = 7.9 Hz, 1 H), 4.26 - 4.20 (m, 1 H), 3.82 - 3.74 (m, 1 H), 3.62 - 3.50 (m, 1 H), 3.24 - 3.13 (m, 1 H), 2.84 - 2.72 (m, 1 H), 2.00 (s, 3H), 1 .97 - 1 .84 (m, 2H), 1 .38 - 1 .13 (m, 2H). LCMS-B: RT 3.134 and 3.196 (two major peaks present) min; m/z 263.2 [M+H]\

(e) 4-(( 1 -Acetylpiperidin-4-yl)amino)-N-(3-((2.3-dihydro- 1 H-inden-2-yl)(2.4-dimethoxy benzyl) amino ) -2-hydroxypropyl)benzamide A59

4-((1 -Acetylpiperidin-4-yl)amino)benzoic acid A58 (0.056 g, 0.213 mmol) was dissolved in DCM (5 mL) and DMF (0.5 mL) and DIPEA (0.068 mL, 0.388 mmol) followed by HATU (0.081 g, 0.213 mmol) were added. The mixture was then stirred at room temperature for 10 min before 1-amino-3-((2.3-dihydro- 1 H-inden-2-yl)(2.4-dimethoxybenzyl)amino)propan-2-ol 112 (0.069 g, 0.194 mmol) in DCM (2 mL) was added. The reaction was then stirred at room temperature for 45 hours. The volatiles were removed in vacuo and the resulting residue diluted with EtOAc (50 mL) followed by sat. aqueous a2CC*3 (50 mL). The layers were separated and the aqueous layer was extracted with EtOAc (50 mL), the combined organic layers were washed with brine (50 mL), dried (Na2S04), filtered and concentrated in vacuo to give the crude product which was purified by silica gel column chromatography (Isolera Biotage, 12 g Si0₂ Cartridge, 0-100% EtOAc in petroleum benzine 40-60 °C, then 0-50% MeOH with 1 % Et₃N in EtOAc) to give the desired compound (0.062 g, 53% yield) as an off- white foam. Ή NMR (400 MHz, d₆-DMSO) δ 7.90 (t, J = 5.4 Hz, 1 H), 7.61 - 7.52 (m, 2H), 7.28 (d, J = 8.3 Hz, 1 H), 7.20 - 7.02 (m, 4H), 6.60 - 6.54 (m, 2H), 6.49 (d, J = 2.4 Hz, 1 H), 6.42 (dd, J = 8.3, 2.4 Hz, 1 H), 6.07 (d, J = 7.9 Hz, 1 H), 4.67 (d, J = 4.2 Hz, 1 H), 4.27 - 4.18 (m, 1 H), 3.81 - 3.66 (m, 9H), 3.60 - 3.46 (m, 3H), 3.30 - 3.24 (m, 1 H), 3.23 - 3.08 (m, 2H), 3.00 - 2.74 (m, 5H), 2.48 - 2.39 (m, 1 H), 2.00 (s, 3H), 1.97 - 1 .82 (m, 2H), 1 .36 - 1 .19 (m, 2H). Missing one proton, possibly obscured behind solvent signal.

(f) 4-((1-Acetylpiperidin-4-yl)amino)-N-(3-((2,3-dihydro-1 H-inden-2-yl)amino)-2-hydroxy propyl) benzamide 64

To 4-((1 -Acetylpiperidin-4-yl)amino)-N-(3-((2, 3-dihydro- 1 H-inden-2-yl)(2.4-dimethoxy benzyl) amino)-2-hydroxypropyl)benzamide A59 (0.060 g, 0.100 mmol) was added TFA (4.0 mL) and the reaction was then stirred at 50 C for 70 hours. The volatiles were removed in vacuo and the resulting residue was diluted with EtOAc (50 mL) followed by 2 M aqueous NaOH (50 mL). The layers were separated and the aqueous layer was extracted with EtOAc (50 mL), the combined organic layers were washed with brine (50 mL), dried (IS^SC ), filtered and concentrated in vacuo to give the desired compound (0.031 g, 69% yield) as an off- white solid.¹H NMR (400 MHz, de-DMSO) δ 8.06 (t, J = 5.6 Hz, 1 H), 7.62 - 7.56 (m, 2H), 7.22-7.15 (m, 2H), 7.14-7.07 (m, 2H), 6.61 -6.54 (m, 2H), 6.08 (d, J= 8.0 Hz, 1H), 4.92 (s, 1H), 4.27-4.17 (m, 1H), 3.83-3.74 (m, 1H), 3.72-3.65 (m, 1H), 3.58-3.48 (m, 2H), 3.30-3.13 (m, 3H, obscured by solvent signal), 3.11 -3.00 (m, 2H), 2.84-2.74 (m, 1H), 2.73 - 2.62 (m, 3H), 2.55 (dd, J= 11.7, 6.8 Hz, 1H), 2.00 (s.3H), 1.97- 1.82 (m, 2H), 1.35 - 1.16 (m, 2H). One NH proton not visible. LCMS-A: RT 4.427 min; m/z 451.3 [M+H]\

Example 43: (S)-A-(3-((2,3 iihydro-1HHnden-2-yl)amino)-2-hydroxypropyl)-4-(1,1- dioxidothiomor holine-4-carbonyl)benzamide 65

(a) (R)-N-(3-((2, 3-Di hydro- 1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino)-2-hydroxypropyl)-4- (1,1 -dioxidothiomorpholine-4-carbonyl)benzamide A60

(f?)-4-((3-((2,3-dihydro-1H-inden-2-yl)(2,4-dimethoxybenzyl)amino)-2- hyd roxy propyl )ca rba moyl )benzoic acid 114 (50 mg, 0.10 mmol), thiomorpholine dioxide (20 mg, 0.15 mmol), MeCN (1 mL), DIPEA (0.052 mL, 0.30 mmol) and HATU (41 mg, 0.11 mmol) were stirred at room temperature. After 2.5 hours methanol (1 mL) was added and the mixture concentrated in vacuo. Chromatography (4 g silica cartridge, 20-100% ethyl acetate/hexanes then 0-10% methanol/ethyl acetate) gave the desired compound as a pale yellow syrup (26 mg, 42% yield). LCMS: RT 3.38 min; m/z (positive ion): 622.4 [M+H]⁺. (b) (S)-N-(3-((2,3-dihydro-1H-inden-2-yl)amino)-2-hydroxypropyl)-4^ 1- dioxidothiomorpholine-4-carbonyl)benzamide 65 (R)-A/-(3-((2,3-Dihydro-1 H-inden-2-yl)(2,4-dimeihoxybenzyl)amino)-2-hyd

dioxidothiomorpholine-4-carbonyl)benzamide A60 (24 mg, 0.042 mmol) and TFA (3 mL) were stirred at 70 C. After four hours the mixture was concentrated in vacuo and the residue dissolved in methanol (3 mL). The mixture was loaded onto a 2g SCX cartridge, the cartridge washed with methanol (25 mL) and eluted with 7M ammonia in methanol (15 mL). The basic eluate was concentrated to give the desired compound as a pale yellow syrup (15 mg, 84% yield). LCMS-B: RT 3.17 min; m/z (positive ion) 472.3 [M+H] \

Example 44: Amide couplings

General Method A

To a stirred solution of the acid (1 eq.) and NEt₃ (3.0 eq.) in dry DCM was added HATU (1.0 eq.). A solution of the amine I6 (1 .0 eq.) in DCM was then added and the reaction stirred at room temperature overnight. The crude mixture was concentrated in vacuo and the residue obtained dissolved in DCM and washed with saturated aqueous

NaHC03. The aqueous layer was further extracted with DCM (x 3) and the combined organic layers were dried (Na2SC>4) then concentrated in vacuo. The crude material was purified by silica gel chromatography (DCM:MeOH = 50:1 ) to yield the desired product.

General Method B

To a solution of the acid (1 .0 eq.) and amine I6 (1 .0 eq.) in DCM were added DIPEA (3.0 eq.), HOBt (0.1 eq.) and EDCI (2.0 eq.). The resulting mixture was stirred at room

temperature overnight. Water was added to the reaction and the mixture extracted with DCM (x 3). The combined organic layers were washed with brine, dried ( a2S0₄) and

concentrated to give the crude product which was purified by column chromatography (DCM: MeOH = 50:1 ) to give the desired product. General Method C

To a solution of the acid (1.0 eq.) in DCM were added DIPEA (3.0 eq.) and HATU (1.0 eq.). The amine 110 (1.0 eq.) in DCM was then added and the solution stirred at room temperature overnight. Water was added and the reaction mixture extracted with DCM x 3. The combined organic layers were washed with brine, dried and concentrated to give the crude product which was purified by column chromatography (DCM: MeOH = 50:1 ) to give the dimethoxybenzyl protected intermediate A.

A solution of the dimethoxybenzyl protected intermediate A (1.0 eq.) in trifluoroacetic acid was stirred at 70 ^°C for 4 h. The mixture was concentrated to give the crude product. A saturated aqueous solution of NaHC03 was then added and the resulting mixture extracted with DCM (x 3). The combined organic layers were dried and concentrated to give the crude product which was purified by preparative TLC (DCM: MeOH = 10:1 ) to give the desired product.

General Method D

To a solution of the acid (1 .0 eq.) and the amine 110 (1 .0 eq.) in DCM were added DIPEA (3.0 eq.), HOBt (0.1 eq.), and EDCI (2.0 eq.). The resulting mixture was stirred at room temperature overnight. Water was added to the reaction mixture and the aqueous extracted with DCM x 3. The combined organic layers were washed with brine, dried ( a2S0₄) and concentrated to give the crude product which was purified by column chromatography (DCM MeOH = 50:1 ) to give the dimethoxybenzyl protected intermediate A.

A solution of the dimethoxybenzyl protected intermediate A (1.0 eq.) in trifluoroacetic acid was stirred at 70 ^°C for 4h. The mixture was concentrated to give the crude product which was suspended in a saturated aqueous solution of NaHC03. The resulting mixture was extracted with DCM (x 3). The organic layers were combined, dried a2S0₄) and

concentrated to give the crude residue which was purified by preparative TLC (DCM: MeOH = 10:1 ) to give the desired product.

General Method E

To a solution of the acid (1 .0 eq.) and amine 118 (1.0 eq.) in DCM were added DIPEA (3.5 eq.), HOBt (1 .4 eq.), and EDCI (1.4 eq.). The resulting mixture was stirred at room

temperature overnight. Water was added to the reaction mixture and the aqueous extracted with DCM x 3. The combined organic layers were washed with brine, dried ( a2S0₄) and concentrated to give the crude product which was purified by column chromatography (DCM: MeOH = 20:1 ) to give the dimethoxybenzyl protected intermediate A.

A solution of the DMB protected intermediate A (1.0 eq.) in trifluoroacetic acid was stirred at 70 ^°C for 2h. The mixture was concentrated to give the crude product which was suspended in a saturated aqueous solution of NaHC0₃ The resulting mixture was extracted with DCM (x 3), the organic layers were combined, dried ( a2S0₄) and concentrated to give the crude residue which was purified by preparative TLC (DCM: MeOH=10:1 ) to give the desired product.

General Method F

To a solution of the acid 114 (1 .0 eq.) in dry DCM were added DIPEA (3.0 eq.) and HATU (1 .0 eq.). The amine (1 .0 eq.) in DCM was then added and the solution stirred at room temperaure overnight. Water was added to the reaction mixture and the aqueous layer extracted with DCM (x 3). The combined organic layers were washed with brine, dried over a2S0₄ and concentrated. The crude residue obtained was purified by column

chromatography (DCM : MeOH = 50:1 ) to give the desired dimethoxybenzyl intermediate A. A solution of compound A (1.0 eq.) in trifluoroacetic acid was stirred at 70 ^°C for 4h. The mixture was concentrated to give the crude product which was suspended in a solution of saturated aqueous NaHC03. The aqueous layer was extracted with DCM (x 3) and the combined organic layers were dried ( a2S0₄) and concentrated. The crude product was purified by preparative TLC (DCM: MeOH = 10:1 ) to give the desired product B General Method G

To a solution of the acid 114 (1.0 eq.) and amine (1 .0 eq.) in DCM were added DIPEA (3.0 eq.), HOBt (0.1 eq.) and EDCI (2.0 eq.).The resulting mixture was stirred at room

temperature overnight. Water was added and the mixture extracted with DCM (x 3). The combined organic layers were washed with brine, dried (Na2SO.₍) and concentrated to give the crude product which was purified by column chromatography (DCM: MeOH = 50:1 ) to give the desired dimethoxybenzyl intermediate A.

A solution of the dimethoxybenzyl intermediate A in trifluoroacetic acid was stirred at 70 ^°C for 4h. The mixture was concentrated to give the crude product which was suspended in a solution of saturated aqueous NaHCC*3. The mixture was extracted with DCM (x 3). The combined organic layers were dried ( a2S0₄) and concentrated to give the crude product which was purified preparative TLC (DCM: MeOH = 10:1 ) to give the desired product B.

General Method H

To a solution of the acid 36 (1.0 eq.) in dry DCM was added Et₃N (5.0 eq.) and HATU (1 .2 eq.). The amine (1.2 eq.) in DCM was then added and the solution was stirred at room temperature overnight. Water was added to the reaction mixture. The mixture was extracted with DCM x 3. The combined organic layers were washed with brine, dried over a2S0₄ and concentrated. The residue was purified by column chromatography (DCM: MeOH = 50:1 ) to give the desired product A. General Method S

To a solution of the acid (1 .0 eq.) and the amine (1.0 eq.) in DCM were added DIPEA (3.0 eq), HOBt (0.1 eq.), and EDCI (2.0 eq.). The resulting mixture was stirred at room

temperature overnight. Water was added to the reaction mixture and the aqueous extracted with DCM (x 3). The combined organic layers were washed with brine, dried ( a2S0₄) and concentrated to give the crude product which was purified by column chromatography (DCM: MeOH = 50:1 ) to give the dimethoxybenzyl protected intermediate A.

A solution of the dimethoxybenzyl protected intermediate A (1.0 eq) in trifluoroacetic acid was stirred at 70 ^°C for 4h. The mixture was concentrated to give the crude product which was suspended in a saturated aqueous solution of NaHC03. The resulting mixture was extracted with DCM (x 3). The organic layers were combined, dried ( a2S0₄) and

concentrated to give the residue which was purified by preparative TLC (DCM: MeOH = 10:1 ) to give the desired product.

General Method J

To a solution of the acid (1 .0 eq) and the amine 11 17 (1.0 eq) in DCM were added DIPEA (3.0 eq), HOBt (0.1 eq) and EDCI (2.0 eq). The resulting mixture was stirred at room temperature overnight. Water was added to the reaction mixture and the aqueous extracted with DCM (x 3). The combined organic layers were washed with brine, dried ( a2S0₄) and concentrated to give the crude product which was purified by preparative TLC (DCM: MeOH =20:1 ) to give the Boc protected intermediate A.

A solution of the Boc protected intermediate A (1 .0 eq) in a saturated solution of HCI in either EtOAc or ΕΪ20 was stirred at room temperature for 3h. The mixture was concentrated to give the crude product. The solid was rinsed with diethyl ether to provide the desired product as a HCI salt. If the product was not pure, the HCI salt was partitioned between DCM and a saturated aqueous solution of NaHCOa and the aqueous extracted with DCM. The combined organic fractions were dried (NazSC ) and concentrated and the crude residue obtained purified by preparative TLC (DCM: MeOH -15:1 v/v) to give the desired product.

General Method K

To a soiution of the acid (1 .0 eq) and the amine 11 17 (1.0 eq) in DCM were added DIPEA (3.0 eq), HOBt (0.1 eq) and EDCI (2.0 eq). The resulting mixture was stirred at room temperature overnight. Water was added to the reaction mixture and the aqueous extracted with DCM (x 3). The combined organic layers were washed with brine, dried ( a2S0₄) and concentrated to give the crude product which was purified by preparative TLC (DCM: MeOH = 20:1 ) to give the Boc protected intermediate A.

A solution of the Boc protected intermediate A (1.0 eq) in DCM/trifluoroacetic acid (8:1 v/v) was stirred at room temperature for 1 h. Water was added and the pH of the solution adjusted to 8-9 by addition of saturated aqueous NaHCC>3 solution. The resulting mixture was extracted with DCM (x 3) and the combined organic fractions were dried ( a2S0₄) and concentrated to give the desired product B.

All Examples in the following Table were prepared using starting materials with the absolute stereochemistry as indicated in the Schemes above, unless stated otherwise in the Method column.

Example 45: V-(2-Amino-3-((2,3-dihydro-1 H-inden-2-yl)amino)propyl)-4-{3-oxa-8- azabic clo[3.2.1]octane-8-carbonyl)benzamide 106

(a) N-(2-Azido-3-((2,3-dihydro- 1H-inden-2-yl)(2A-dimethoxybenzyl)amino)propyl)-4-(3-oxa- 8-azabicyclo[3.2.1 ]octane-8-carbonyl)benzamide A62

To a solution of 4-(3-oxa-8-azabicyclo[3.2.1 ]octane-8-carbonyl)-/V-(3-((2,3-dihydro-1 Z+inden- 2-yl)(2.4-dimethoxybenzyl)amino)-2-hydroxypropyl)benzamide (Racemic analogue of Intermediate A in synthesis of 103, made in analogous way but with a racemic starting material) (100 mg, 0.17 mmol) in THF (5 mL) was added DPPA (95 mg, 0.34 mmol) and DBU (52 mg, 0.34 mmol). The resulting mixture was stirred at 60 C overnight. LCMS showed the starting materials remained, further DPPA (46 mg, 0.17 mmol) and DBU (25 mg, 0.17 mmol) were added and the mixture was stirred at 60 C overnight. The solvent was removed and the residue was purified by column chromatography (0-25% EtOAc/petroleum ether) to give the desired compound as an off-white solid (85 mg, 82%): ¹H NMR (400 MHz, CDC ) δ 7.66-7.64 (m, 2H), 7.56-7.54 (m, 2H), 7.21 -7.14 (m, 6H), 6.45-6.38 (m, 2H), 4.75 (br s, 1 H), 3.95-3.83 (m, 5H), 3.79-3.76 (m, 5H), 3.74-3.69 (m, 6H), 3.65-3.59 (m, 2 H), 3.35- 3.30 (m, 1 H), 3.21-3.12 (m, 4H), 2.07-2.05 (m, 4 H): LCMS: RT 2.65 min, m/z 625.3 [M+H]'

(b) N-(2-Amino-3-((2,3-dihydro- 1H-inden-2-yl)(2,4-dimethoxybenzyl)amino)propyl)-4-(3-oxa- 8-azabicyclo[3.2. 1 ]octane-8-carbonyl)benzamide A63

To a solution of A/-(2-azido-3-((2,3-dihydro-1 H-inden-2-yl)(2,4- dimethoxybenzyl)amino)propyl)-4-(3-oxa-8-azabicyclo[3.2.1]octane-8-carbonyl)benzamide A62 (70 mg, 0.1 1 mmol) in THF (2 mL) was added PPh₃ (44 mg, 0.17 mmol). The resulting mixture was stirred at 50 °C for 3 h and then water (0.5 mL) was added dropwise and the mixture stirred at 30 °C overnight. The solvent was removed and the residue diluted with DCM (20 mL). The organic layer was dried (Na2S0₄) and concentrated to give the crude product as a pale yellow solid (100 mg). The product was used directly for the next step without further purification. LCMS: RT 2.60 min, m/z 599.3 [M+H] '

(c) N-(2-Amino-3-((2, 3-dihydro- 1 H-inden-2-yl)amino)propyl)-4-(3-oxa-8- azabicyclo[3.2. 1 ]octane-8-carbonyl)benzamide 106

A mixture of A/-(2-amino-3-((2.3-dihydro-1 /-/-inden-2-yl)(2,4-dimethoxybenzyl)amino)propyl)- 4-(3-oxa-8 azabicyclo[3.2.1 ]octane-8-carbonyl)benzamide A63 (67 mg, 0.1 1 mmol) in TFA (1 mL) was stirred at 70 °C for 6 h. The solvent was removed and the residue was diluted with water (5 mL). The aqueous layer was washed with DCM (2 x 10 mL) and the pH adjusted to 9 by addition of solid a2CC»3. The aqueous layer was extracted with (10% MeOH in CH2CI2) (6x10 mL). The combined organic layers were dried (Na2SO.i) and concentrated to give the desired compound as a yellow solid (24 mg, 48%): ¹ H NMR (400 MHz, dt-MeOD) δ 7.91- 7.89 (m, 2H), 7.61-7.59 (m, 2H), 7.17-7.08 (m, 4H), 4.66 (br s, 1 H), 3.95 (s, 1 H), 3.83-3.69 (m, 4H), 3.61-3.56 (m, 2H), 3.49-3.45 (m, 1 H), 3.24-3.15 (m, 2H), 2.98-2.93 (m, 1 H), 2.83- 2.62 (m, 4H), 2.07-2.01 (m, 4H): LCMS:RT 0.61 min, m/z 449.2 [M+H]^* Example 46: W-(3-Amino-2-(((2,3-dihydro-1 «-inden-2-yl)amino)methyl)-3-oxopropyl)-4- (mor holine-4-carbonyl)benzamide 107

(a) Methyl 2-(( 1 , 3-dioxoisoindolin-2-yl)methyl)acrylate A63

To a solution of methyl 2-(bromomethyl)acrylate (2.0 g, 1 1 .2 mmol) in toluene (50 mL) was added potassium 1 ,3-dioxoisoindolin-2-ide (2.0 g, 1 1 .2 mmol) and 18-crown-6 (2.9 g, 1 1.2 mmol). The resulting mixture was stirred at 90 °C overnight. The reaction was partitioned with water (50 mL) and the aqueous layer further extracted with EtOAc (2x50 mL). The combined organic layers were washed with water (4x100 mL) and brine (100 mL), dried (Na2SO.i) and concentrated to give the desired compound as a white solid (2.3 g, 85%): LCMS:RT 2.20 min, m/z 246.1 [M+H]⁺ (b) Methyl 3-((2.3-dihydro- 1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino)-2-((1 ,3- dioxoisoindolin-2-yl)methyl)propanoate A64

To a solution of methyl 2-((1 ,3-dioxoisoindolin-2-yl)methyl)acrylate A63 (2.0 g, 8.2 mmol) in CHC (15 mL) was added A/-(2.4-dimethoxybenzyl)-2,3-dihydro-1 H-inden-2-amine I9 (2.3 g, 8.2 mmol). The resulting mixture was heated at reflux for 3 days. The solvent was removed and the residue purified by column chromatography (0-17% Petroleum ether/EtOAc) to give the desired compound as a yellow oil (2.6 g, 60%): Ή NMR (400 MHz, CDC ) δ 7.84-7.82 (m, 2H), 7.72-7.70 (m, 2H), 7.29-7.27 (m, 1 H), 7.16-7.15 (m, 2H), 7.1 1 -7.08 (m, 2H), 6.47- 6.44 (m, 1 H), 6.42-6.40 (m, 1 H), 3.93-3.92 (m, 1 H), 3.84-3.81 (m, 3H), 3.78-3.77 (m, 6H), 3.76-3.73 (m, 2 H), 3.62 (s, 3H), 3.16-3.13 (m, 1 H), 3.00-2.93 (m, 4H), 2.69-2.64 (m, 1 H): LCMS:RT 2.41 min, m/z 529.3 [M+H]'

(c) Methyl 3-amino-2-(((2,3-dihydro- 1H-inden-2-yl)(2.4- dimethoxybenzyl)amino )methyl)propanoate A 65

To a solution of methyl-3-((2.3-dihydro-1 f -inden-2-yl)(2,4-dimethoxybenzyl)amino)-2-((1 _!3- dioxoisoindolin-2-yl)methyl)propanoate A64 (2.6 g, 4.9 mmol) in MeOH (30 mL) was added DCM (5 mL) and hydrazine hydrate (85% aqueous solution, 270 mg, 5.4 mmol). The resultant mixture was stirred at room temperature overnight. The precipitate which formed was removed by filtration and the filtrate was concentrated in vacuo. The residue was diluted with DCM (100 mL) and a 2 M aqueous HCI solution (50 mL) was added. The pH of the aqueous layer was adjusted to 10 by addition of 6 M aqueous NaOH solution and extracted with DCM (3x100 mL). The organic layers were combined, dried (Na2SC> ), filtered and concentrated in vacuo to give the desired compound as a yellow oil (1 .0 g, 51 %). The compound was used in the next step without further purification: LCMS:RT 1.38 min, m/z 399.3 [M+H]'

(d) Methyl 3-((2,3-dihydro- 1H-inden-2-yl)(2A-dimethoxybenzyl)amino)-2-((4-(m

carbonyl)benzamido)methyl)propanoate A66

To a solution of 4-(morpholine-4-carbonyl)benzoic acid I27 (420 mg, 1 .80 mmol) in DCM (15 mL) was added HATU (685 mg, 1.80 mmol) and DIPEA (526 mg, 4.07 mmol) and the mixture was stirred at room temperature for 30 min. Methyl 3-amino-2-(((2.3-dihydro-1 H- inden-2-yl)(2,4-dimethoxybenzyl)amino) methyl) propanoate A65 (650 mg, 1.63 mmol) was added and the mixture was stirred at room temperature overnight. The mixture was diluted with DCM (70 mL) washed with water (70 mL), saturated aqueous NaHCCXi (70 mL), water (70 mL) and brine (70 mL), dried (Na2SO.i) and concentrated. The residue was purified by column chromatography (0-30% EtOAc in DCM) to give the desired compound as a light solid (450 mg, 45%): ¹H NMR (400 MHz, CDC ) δ 8.23 (br s, 1 H), 7.68-7.66 (m, 2H), 7.34- 7.33 (m, 2H), 7.19-7.17 (m. 2H), 7.12-7.10 (m, 3H), 6.43-6.41 (m, 1 H), 6.34 (s, 1 H), 3.99- 3.84 (m, 4H), 3.82-3.70 (m, 12H), 3.56 (s, 3H), 3.52-3.38 (m, 4H), 3.04 (br s, 3H), 2.97-2.94 (m, 3H): LCMS:RT 2.21 min, m/z 616.4 [M+H]⁺

(e) 3-((2,3-Dihydro-1H-inden-2-yl)(2,4-dimethoxybenzyl)amino)-2-((4-(morpholine-4- carbonyl)benzamido)methyl)propanoic acid A67

To a solution of methyl 3-((2,3-dihydro-1 /+inden-2-yl)(2.4-dimethoxybenzyl)amino)-2-((4- (morpholine-4-carbonyl)benzamido)methyl)propanoate A66 (100 mg, 0.16 mmol) in MeOH (5 mL) was added a solution of NaOH (13 mg, 0.32 mmol) in water (0.5 mL). The resulting mixture was stirred at room temperature overnight. The solvent was removed and the residue obtained dissolved in water. The pH of the aqueous solution was adjusted to 3 by addition of a 2 M aqueous HCI solution. The aqueous layer was extracted with DCM (3x20 mL) and the combined organic layers dried ( a2S0₄) and concentrated to give the desired compound as a yellow solid (100 mg, 100%): LCMS: RT 2.38 min, m/z 602.4 [M+H]^*

(f) N-(3-Amino-2-(((2, 3-dihydro- 1 H-inden-2-yl)(2, 4-dimethoxybenzyl)amino)methyl)-3- oxopropyl) -4-(morpholine-4 -carbonyl)benzamide A 68

To a solution of 3-((2.3-dihydro-1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino)-2-((4-

(morpholine-4-carbonyl)benzamido)methyl)propanoic acid A67 (50 mg, 0.08 mmol) in DMF (0.5 mL) was added HATU (32 mg, 0.08 mmol) and DIPEA (32 mg, 0.24 mmol). The mixture was stirred at room temperature for 30 min, then poured into aqueous ammonia (26%, 2 mL) and stirred at room temperature for 1 h. The mixture was diluted with DCM (50 mL) and washed with water (2x50 mL) and brine (20 mL), dried ( a?S04) and concentrated. The residue was purified by preparative TLC (10% methanol in DCM) to give the desired compound as a yellow solid (30 mg, 40%): LCMS: RT 2.18 min, m/z 601.3 [M+H]'

(g) N-(3-Amino-2-(((2,3-dihydro- 1H-inden-2-yl)amino)methyl)-3-oxopropyl)-4-(morpholine-4- carbonyl)benzamide 107

A mixture of A/-(3-amino-2-(((2.3-dihydro-1 /-/-inden-2-yl)(2,4-dimethoxybenzyl)amino)methyl)- 3-oxopropyl)-4-(morpholine-4-carbonyl)benzamide A68 (30 mg, 0.05 mmol)in TFA (1 mL) was stirred at 70 °C for 6 h. The solvent was removed and the residue suspended in water. The pH was adjusted to 8 by addition of saturated aqueous NaHCOa solution. The aqueous layer was extracted with DCM (2 x 20 mL) and the combined organic layers washed with water (20 mL) and brine (20 mL), dried ( a2S0₄) and concentrated. The residue obtained was purified by preparative TLC (10% methanol in DCM) to give the desired compound as a yellow solid (10 mg, 45%): Ή NMR (400 MHz, 4-methanol) δ 7.94-7.92 (m, 2H), 7.56-7.52 (m 2H), 7.25-7.16 (m, 4H), 3.96-3.89 (m, 1 H), 3.77-3.61 (m, 8H), 3.42 (brs, 2H), 3.37-3.31 (m, 2H), 3.27-3.22 (m, 1 H), 3.13-3.01 (m, 4H): LCMS:RT 3.02 min, m/z 451 .2 [M+H]'

Example 47: yV-(3-((2,3-Dihydro-1 H-inden-2-yl)amino)-2-(hydroxymethyl)propyl)-4- (mor holine-4-carbonyl)benzamide 108

(a) 3-Amino-2-(((2.3-dihydro- 1 H-inden-2-yl)(2A-dimethoxybenzyl)amino)methy^

A69

To a solution of methyl 3-amino-2-(((2,3-dihydro-1 /+inden-2-yl)(2.4-dimethoxybenzyl)amino) methyl)propanoate A65 (50 mg, 0.13 mmol) in THF (4 mL) was added LiAIH₄ (10 mg, 0.26 mmol). The resulting mixture was stirred at room temperature for 2 h. The mixture was quenched with water (2 x 0.5 mL) 10% aqueous NaOH (0.5 mL) and water (0.5 mL) and stirred at room temperature for 10 min. The solvent was removed, water was added and the mixture was extracted with DCM (3 x10 mL). The combined organic layers were dried (Na2SO.t) and concentrated to give the desired compound as yellow oil (45 mg, 98%):

LCMS: RT 0.96 min, m/z 371.2 [M+H]' (b) N-(3-((2,3-Dihydro- 1H-inden-2-yl)(2A-dimethoxybenzyl)amino)-2-^

4-(morpholine-4-carbonyl)benzamide A 70

To a solution of 3-amino-2-(((2,3-dihydro-1 H-inden-2-yl)(2,4-dimethoxybenzyl)

amino)methyl) propan-1-ol A69 (180 mg, 0.48 mmol) in DCM (10 mL) was added 4- (morpholine-4-carbonyl)benzoic acid I27 (125 mg, 0.53 mmol), DIPEA (186 mg, 1.44 mmol), HOBt (7 mg, 0.05 mmol) and EDCI (1 10 mg, 0.58 mmol). The resulting mixture was stirred at room temperature for two days. The mixture was diluted with DCM (50 mL) and the organic layer washed with water (50 mL), saturated aqueous NaHCC (50 mL), water (50 mL) and brine (50 mL), dried Na2S0₄) and concentrated. The residue was purified by column chromatography (0-2% MeOH in DCM) to give the desired compound as a yellow solid (80 mg, 28%): LCMS: RT 2.25 min, m/z 588.2 [M+H]⁺ (c) N-(3-((2,3-Dihydro-1H-inden-2-yl)amino)-2-(hydroxymethyl)propyl)-4-(morph

carbony!)benzamide 108

A mixture of A/-(3-((2,3-dihydro-1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino)-2-(hydroxyl methyl) propyl)-4-(morpholine-4-carbonyl)benzamide A70 (80 mg, 0.14 mmol) in TFA (4 mL) was stirred at 70 C for 6 h. The solvent was removed and the residue suspended in water. The pH was adjusted to 8 by addition of saturated aqueous NaHCOa solution. The aqueous layer was extracted with DCM (3 x 20 mL) and the combined organic layers washed with water (40 mL) and brine (40 mL), dried (Na2SO.i) and concentrated. The residue obtained was purified by preparative TLC (10% methanol in DCM) to give the desired compound as a yellow solid (30 mg, 51 %): ¹H NMR (400 MHz, Methanol-d.,) δ 7.92-7.90 (m, 2 H), 7.54-7.52 (m, 2 H), 7.22-7.17 (m, 4 H), 3.87-3.42 (m, 14 H), 3.31 (m, 1 H, overlap), 3.02-2.89 (m, 4 H), 2.19-2.15(m, 1 H). LCMS: RT 3.13 min, m/z 438.2 [M+H] '

Example 48: (S)-/V-(3-((2,3-Dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)-2-((1 - (methylsulfonyl)azetidin-3-yl)amino)isonicotinamide bis 2,2,2-trifluoroacetic acid salt 109

(a) Methyl 2-(( 1 -(tert-butoxycarbonyl)azetidin-3-yl)amino)isonicotinate A71

To a solution of methyl 2-bromoisonicotinate (3.0 g, 14.0 mmol) in dioxane (40mL) was added ie/t-butyl 3-aminoazetidine-1 -carboxylate (3.6 g, 21.0 mmol), cesium carbonate (9.1 g, 28.0 mmol), Xantphos (1 .6g, 2.8 mmol) and Pd?(dba)3 (1.3 g, 1 .4 mmol) under a nitrogen atmosphere. The reaction was heated at reflux overnight, then concentrated and the residue diluted with water (40 mL). The resulting mixture was extracted with DCM (3 x 40 mL) and the combined organic extracts washed with saturated aqueous NaHCOa (3 x 30 mL) and brine (3 x 30 mL), dried ( a2S0₄) and concentrated to give a crude residue which was purified by column chromatography (20% ethyl acetate /petroleum ether) to give the desired compound as a white solid (940mg, 22%). H NMR (400MHz, CDC ) δ 8.20-8.19 (m, 1 H), 7.15-7.14 (m, 1 H), 6.93 (s, 1 H), 5.16-5.15 (m, 1 H), 4.59-4.51 (m, 1 H), 4.35-4.31 (m, 2H), 3.91 (s, 3H), 3.78-3.75 (m, 2H), 1.44 (s,9H); LCMS RT 2.45 min; m/z 308.1 [M+H]\

(b) 2-((1-(tert-Butoxycarbonyl)azetidin-3-yl)amino)isonicotinic acid A72

To a solution of methyl 2-((1-(iert-butoxycarbonyl)azetidin-3-yl)amino)isonicotinate A71 (930 mg, 3.1 mmol) in a mixture of THF (10 mL), MeOH (5 mL) and water (4 mL) was added lithium hydroxide hydrate (1 .3 g, 30.3 mmol). The resulting mixture was stirred at room temperature overnight. The mixture was concentrated under reduced pressure, then diluted with water. The pH of the aqueous solution was adjusted to 1 -2 by addition of 4M aqueous HCI. Removal of the solvent provided the crude product containing lithium chloride (1 .3g). The material was carried forward without further purification. LCMS RT 1.26 min; m/z 294.1 [M+H]⁺

(c) (R)-ten-Butyi-3-((4-((3-((2.3-dihydro- 1H-inden-2-yl)(2A-dimethoxy

hydroxypropyl)carbamoyl)pyridin-2-yl)amino)azetidine- 1 -carboxylate A73

To a suspension of 2-((1-(iert-butoxycarbonyl)azetidin-3-yl)amino)isonicotinic acid A72 (600 mg,2.1 mmol) in DCM (10 mL) was added EDCI (786 mg, 4.1 mmol), HOBt (555mg, 4.1 mmol) and DIEA (1.3 g, 10.3 mmol). A solution of (fl)-1-amino-3- ((2,3-dihydro-1 H-inden-2- yl)(2,4-dimeth-oxybenzyl)amino)propan-2-ol 110 (875 mg, 2.5 mmol) in DCM (10 mL) was added and the resulting mixture stirred at room temperature overnight. The mixture was diluted with water (20 mL) and DCM (20 mL) and the aqueous layer extracted with DCM (3 x 10 mL). The combined organic extracts were washed with saturated aqueous NaHCC (3 x 10mL) and brine (3 x 10mL), dried (Na2S0₄) and concentrated. The residue obtained was purified by silica gel column chromatography (ethyl acetate: petroleum ether=2:1 ) to give the desired compound as an off-white solid (600 mg, 46%). LCMS RT 2.42 min; m/z 632.4

[M+H]⁺ (d) (R)-2-(Azetidin-3-ylamino)-N-(3-((2,3-dihydro-1H-inden^

dimethoxybenzyl)amino)-2-hydroxypropyl)isonicotinamide A74

A solution of (fl)-terf-butyl 3-((4-((3-((2,3-dihydro-1 H-inden-2-yl)(2,4-dimethoxybenzyl) amino)-2-hydroxypropyl)carbamoyl)pyridin-2-yl)amino)azetidine-1 -carboxylate A73 (600 mg, 0.95 mmol) in a saturated solution of HCI (g) in EtOAc (30 mL) was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure and the residue obtained dissolved in DCM (30 mL) and partitioned with water. The aqueous layer neutralized by addition of saturated aqueous aHCOa to a pH of 7-8. The organic layer was separated and the aqueous solution further extracted with DCM (3 x 10 mL). The combined organic extracts were washed with saturated brine (3 x 10 mL), dried (Na2S0₄) and concentrated under reduced pressure to give the desired product as a yellow solid (400 mg, 79%) which was used without further purification. LCMS RT 3.13 min; m/z 532.3 [M+H]⁺

(e) (R)-N-(3-((2,3-Dihydro-1H-inden-2-yl)(2A-dimethoxybenzyl)amino)-2-hyd

((1 -(methylsulfonyl)azetidin-3-yl)amino)isonicotinamide A75

To a solution of (f?)-2-(azetidin-3-ylamino)-W-(3-((2,3-dihydro-1 H-inden-2-yl) (2,4- dimethoxybenzyl)amino)-2-hydroxypropyl)isonicotinamide A74 (150 mg, 0.28 mmol) in DCM (5 mL) was added triethylamine (85 mg, 0.84 mmol). A solution of methanesulfonyl chloride (33 mg, 0.28 mmol) in DCM (1 mL) was added dropwise into the mixture and the reaction stirred at room temperature for 2h. Water (15 mL) and DCM (10 mL) were added to the mixture and the aqueous layer extracted with DCM (3 x 5 mL). The combined organic extracts were washed with brine (3 x 10 mL), dried ( a2S0₄) and concentrated. The residue was purified by preparativeTLC (5% methanol in dichloromethane) to give the desired compound as a yellow solid (70 mg, 41 %). LCMS RT 2.01 min; m/z 610.3 [M+H] '

(f) (S)-N-(3-((2,3-Dihydro- 1 H-inden-2-yl)amino)-2-hydroxypropyl)-2-((1- (methylsulfonyl)azetidin-3-yl)amino)isonicotinamide bis 2, 2, 2-trifluoroacetic acid salt 109 A solution of (f?)-A/-(3-((2,3-dihydro-1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino) -2- hydroxypropyl)-2-((1-(methylsulfonyl)azetidin-3-yl)amino)isonicotinamide A75 (60 mg, 0.1 mmol) in trifluoroacetic acid (2 mL) was heated at reflux overnight. The mixture was concentrated under reduced pressure and the residue dissolved in DCM (10 mL) and partitioned with saturated aqueous NaHCO.3 (10 mL). The aqueous layer was further extracted with DCM (3 x 5 mL) and the aqueous layer was concentrated under reduced pressure to give a crude residue. The residue was suspended in methanol/dichloromethane (1/20) (40 mL) and the salts removed by filtration. The organic solvent was removed to give the crude product which was purified by preparativeHPLC to give the bis-TFA salt of the desired compound (23 mg, 51 %) as an off-white solid. ¹ H NMR (400 MHz, MeOH-d₄) δ 8.20- 8.18 (m, 1 H), 7.40 (m, 1 H), 7.27-7.20 (m, 5H), 4.87-4.81 (m, 1 H), 4.68-4.63 (m, 1 H), 4.58- 4.52 (m, 1 H), 4.15-4.08 (m, 2H), 3.56-3.52 (m, 1 H), 3.48-3.36 (m, 5H), 3.28-3.24 (m, 1 H), 3.19-3.1 1 (m, 2H), 3.06-3.01 (m, 1 H), 2.97 (s, 3H); LCMS RT 0.26 min; m/z 460.2 [M+H]⁺ (freebase)

Example 49: (S)-2-((1 -Acetylazetidin-3-yl)amino)-/V-(3-{(2,3-dihydro-1 H-inden-2- l)amino)-2-hydroxypropyl)isonicotinamide bis 2,2,2-trif!uoroacetic acid salt 110

(a) (R)-2-((1-acetylazetidin-3-yl)amino)-N-(3 (2,3-dihydro- 1H-i

dimethoxybenzyl)amino)-2-hydroxypropyl)isonicotinamide A76

To a solution of (f?)-2-(azetidin-3-ylamino)-/V-(3-((2,3-dihydro-1 H-inden-2-yl) (2.4- dimethoxybenzyl)amino)-2-hydroxypropyl)isonicotinamide A74 (220 mg, 0.41 mmol) in DCM (5 mL) was added triethylamine (212 mg, 2.10 mmol) followed by a solution of acetic anhydride (42 mg, 0.41 mmol) in DCM (1 mL) dropwise. The resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated and the residue dissolved in DCM (2 OmL) and partitioned with water (15 mL). The aqueous layer was extracted with DCM (3 x 10 mL) and the combined organic extracts washed with brine (3 x 10 mL), dried ( a2S0₄) and concentrated. The residue obtained was purified by preparativeTLC (5% methanol in dichloromethane) to give the desired compound as a white solid (150 mg, 64%). LCMS RT 2.04 min; m/z 574.3 [M+H]\

(b) (S)-2-((1 -Acetylazetidin-3-yl)amino)-N-(3-((2,3-dihydro- 1H-inden-2-yl

hydroxypropyl)isonicotinamide bis 2, 2, 2-trifluoroacetic acid salt 110

A solution of ( )-2-((1-acetylazetidin-3-yl)amino)-A/-(3-((2,3-dihydro-1 H-inden-2-yl) (2,4- dimethoxybenzyl)amino)-2-hydroxypropyl)isonicotinamide A76 (130 mg, 0.23 mmol) in trifluoroacetic acid (4 mL) was heated at reflux overnight. The mixture was concentrated under reduced pressure and the residue obtained dissolved in DCM (10 mL) and partitioned with saturated aqueous NaHCO.3 (10 mL). The aqueous layer was washed with DCM (3 x 5 ml_) then concentrated to give a crude residue. The residue obtained was suspended in a mixture of methanol/dichloromethane (1/20) (40 mL) and the inorganic salts removed by filtration. The filtrate was concentrated and the residue was purified by preparativeHPLC to give the bis-TFA salt of the desired compound (40 mg, 41 %) as an off-white solid: ¹ H NMR (400 MHz, MeOD) δ 8.08-8.06 (m, 1 H), 7.28-7.20 (m, 5H), 7.16-7.14 (m, 1 H), 4.66-4.60 (m, 2H), 4.42-4.38 (m, 1 H), 4.16-4.10 (m, 3H), 3.93-3.90 (m, 1 H), 3.57-3.53 (m, 1 H), 3.48-3.39 (m, 3H), 3.28-3.24 (m, 1 H), 3.18-3.1 1 (m, 2H), 3.07-3.01 (m, 1 H), 1 .90 (s, 3H); LCMS: RT 2.37 min; m/z 424.3 [M+H]\

Example 50: (S)-A -(3-((2,3-Dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)-A -methyl-4- (mor holine-4-carbonyl)benzamide 111

(a) (R)-Tert-butyl (3-((2,3-dihydro- 1H-inden-2-yi)(2,4-dimethoxybenzyl)amino)-2- hydroxypropyl) carbamate A 77

To a solution of (ft)-1-amino-3-((2,3-dihydro-1 H-inden-2-yl) (2, 4-dimethoxybenzyl) amino)propan-2-ol 110 (500 mg, 1 .4 mmol) in dichloromethane (15 mL) was added triethylamine (284.0 mg, 2.8 mmol) and (Βο Ο (305.0 mg, 1 .4 mmol). The resulting mixture was stirred at room temperature overnight. Water was added to the solution and the mixture was extracted with dichloromethane (20 mL x 3). The organic layer was dried ( a2S0₄) and concentrated to give the crude product which was purified by column chromatography (Petroleum ether: ethyl acetate=3:1 ) to give the desired product (380 mg, 59%) as yellow oil. LCMS: RT 2.21 min; m/z 457.3 [M+H] ' (b) (R)- 1 -((2, 3-Dihydro- 1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino)-3-(methylamino)propan- 2-OI A78 To a solution of (f?)-£erf-butyl (3-((2,3-dihydro-1 H-inden-2-yl) (2.4-dimethoxybenzyl)amino)-2- hyd roxypropyl )ca rba mate A77 (670 mg, 1 .47 mmol) in dry THF (10 mL) was added lithium aluminum hydride (334.3 mg, 8.82 mmol). The mixture was heated at reflux for 10 h. To the mixture was added a solution of NaOH (1.05 g, 26.4mmol) in water (1 mL) and the precipitate that formed removed by filtration. The filter cake was rinsed with THF (200 mL x 3) and the filtrate combined, dried ( a2S0₄) and concentrated to give the crude product which was purified by column chromatography (Petroleum ether: ethyl acetate=1 :1 to dichloromethane : methanol =20:1 ) to give the desired product (230 mg, 42%) as yellow oil. LCMS: RT 0.37 min; m/z 371.3 [M+H]⁺

(c) (S)-N-(3-((2,3-Dihydro- 1H-inden-2-yl)(2A-dimethoxybenzyl)ami

methyl-4-(morpholine-4-carbonyl)benzamide A 79

To a solution of I27 (127.0 mg, 0.54mmol) and (f?)-1 -((2,3-dihydro-1 H-inden-2-yl)(2,4- dimethoxybenzyl)amino)-3-(methylamino)propan-2-ol A78 (200 mg, 0.54mmol) in DCM (910 mL) was added DIPEA (279.0 mg, 2.16 mmol), HOBt (1 14.8 mg, 0.75 mmol), and EDCI (143.0 mg, 0.75 mmol). The resulting mixture was stirred at room temperature overnight. Water was added to the reaction mixture and the mixture extracted with DCM. The combined organic layers were washed with brine, dried (NazSC ) and concentrated to give the crude product which was purified by column chromatography (DCM: MeOH = 50:1 ) to give the desired product (170 mg, 53%) as yellow oil. LCMS: RT 2.10 min, m/z 588.3 [M+H]⁺

(d) (S)-N-(3-((2,3-Dihydro- 1H-inden-2-yl)amino)-2-hydroxypro

carbonyl)benzamide 11 1

A mixture of (S)- V-(3-((2,3-dihydro-1 H-inden-2-yl)(2.4-dimethoxybenzyl)amino)-2- hydroxypropyl)-/V-methyl-4-(morpholine-4-carbonyl)benzamide A79 (170 mg, 0.29 mmol) in TFA (5 mL) was stirred at 70 ' C for 6 h. The solvent was removed and the residue suspended in water. The pH was adjusted to 8 by addition of saturated aqueous NaHC03 solution. The aqueous layer was extracted with DCM (2 x 20 mL) and the combined organic layers washed with water (20 mL) and brine (20 mL), dried (Na2SO.i) and concentrated. The residue obtained was purified by preparativeTLC (10% methanol in DCM) to give the desired compound as a white solid (40 mg, 30%): Ή NMR (400 MHz, Methanol-d4) δ 7.57- 7.55 (m, 4 H), 7.26 - 7.21 (m, 4 H), 4.27 (m, 1 H), 4.08 -4.04 (m, 1 H), 3.76 -3.76 (m, 5 H), 3.64 - 3.57 (m, 2 H), 3.58-3.56 (m, 1 H) 3.55-3.35 (m, 4 H), 3.22-3.05 (m, 5 H) 3.00-2.86 (m, 2 H), LCMS: RT 0.49 min, m/z 438.2 [M+H]⁺ Example 51 : Alternative synthesis of 4-(3-Oxa-8-azabicyclo[3.2.1]octane-8-carbonyi)- A - S)-3-{(2,3 lihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)-2-ethoxybenzamide 77

(a) 4-(3-Oxa-8-azabicyclo[3.2.1]octane-8-carbonyl)-N-((R)-3-((2,3-dihydro- 1H-inden-2- yl)(2,4-dimethoxybenzyl)amino)-2-hydroxypropyl)-2-ethoxybenzamide A5

To a solution of (f?)-4-((3-((2,3-dihydro-1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino)-2- hydroxypropyl)carbamoyl)-3-ethoxybenzoic acid I25 (5.2 g, 9.48 mmol) and 3-oxa-8- azabicyclo[3.2.1 ]octane hydrochloride (1.42 g,9.48 mmol) in DCM (20 mL) was added DIPEA (4.8 g, 37.9 mmol), HOBt (0.29 g,1 .9 mmol), and EDCI (3.6 g, 19.0 mmol). The resulting mixture was stirred at room temperature overnight. Water (20 mL) was added to the reaction mixture and the aqueous extracted with DCM (20 mL x 3). The combined organic layers were washed with brine, dried (Na2SO.t) and concentrated to give the crude product which was purified by column chromatography (DCM: MeOH = 100:1 ) to give the desired product (5.8 g, 95%) as yellow oil. LCMS: RT 2.22 min. m/z 644.3 [M+H]⁺

(b) Alternate synthesis of 4-(3-Oxa-8-azabicyclo[3.2. 1]octane-8-carbonyl)-N-((S)-3-((2,3- dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)-2-ethoxybenzamide 77

A solution of 4-(3-oxa-8-azabicyclo[3.2.1 ]octane-8-carbonyl)-/V-(( ?)-3-((2,3-dihydro-1 H- inden-2-yl)(2,4-dimethoxybenzyl)amino)-2-hydroxypropyl)-2-ethoxybenzamide A5 (5.8 g, 90.0 mmol) in trifluoroacetic acid (30 mL) was stirred at 70 C for 4 h. The mixture was concentrated to give the crude product which was suspended in saturated aqueous NaHC0₃ (100 mL). The resulting mixture was extracted with DCM (x 3) and the organic layers were combined, dried ( a2S0₄) and concentrated to give a crude residue which was purified by column chromatography (DCM: MeOH = 80:1 >20:1 ) to give the desired product (2.13 g, 48%) as a white solid. LCMS: RT 1.87 min, m/z 494.3 [M+H]⁺. ¹H NMR (400 MHz, Methanol- d4) δ 8.03-8.01 (m, 1 H), 7.24 (brs, 1 H), 7.18-7.15 (m, 3 H), 7.13-7.09 (m, 2 H), 4.64 (s, 1 H), 4.30-4.25 (m, 2 H), 3.97-3.92 (m, 2 H), 3.84-3.71 (m, 3 H), 3.67-3.57 (m, 3 H), 3.49-3.44 (m, 1 H), 3.23-3.16 (m, 2 H), 2.85-2.71 (m, 4 H), 2.06-2.00 (m, 4 H), 1 .54-1.51 (m, 3 H).

Example 52: (H)-A?-(3-((2,3-Dihydro-1 H-inden-2-yl)(methyl)amino)-2-hydroxypropyl)-4- morpholine-4-carbonyf)benzamide 112

(a) (R)-N-(3-((2,3-dihydro- 1H-inden-2-yl)(2A-dimethoxybenzyl)amino)-2-hydroxypropyl)-4- (morpholine-4-carbonyl)benzamide A80

To a solution of 4-(morpholine-4-carbonyl)benzoic acid I27 (659 mg, 2.8 mmol) in DCM (35 mL) at 0 C was added DIPEA (1 .09 g, 8.4 mmol) and HATU (1.17 g. 3.08 mmol). The mixture was stirred at 0 °C for 30 min. To the mixture was added a solution of (f?)-1 -amino-3- ((2,3-dihydro-1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino)propan-2-ol 110 (1.0 g, 2.8 mmol) in DCM (15 mL). The resulting mixture was allowed to warm to room temperature and stirred overnight. Water was added to the mixture and the solution extracted with DCM (30 mL x 3). The combined organic layers were dried and concentrated to give the crude product which was purified by column chromatography (DCM:MeOH=50:1 ) to give the desired product (1 .38 g, about 70% purity) as a yellow oil. LCMS: RT 2.00 min; m/z 574.3 [M+H]'

(b) N-(2,4-dimethoxybenzyl)-N-((R)-2-hydroxy-3-(4-(morpholine-4- carbonyl)benzamido )propyl) -N-methyl-2,3-dihydro- 1 H-inden-2-aminium iodide A81

To a solution of (f?)-/V-(3-((2,3-dihydro-1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino)-2- hydroxypropyl)-4-(morpholine-4-carbonyl)benzamide A80 (400 mg, 0.68mmol) in DMF (5 mL) was added K₂C0₃ (282 mg, 2.04 mmol) and iodomethane (288 mg, 2.04 mmol). The resulting mixture was stirred at 55 °C in a sealed tube overnight. Water was added to the mixture and the solution extracted with DCM (20 mL x3). The organic layer was washed with brine (20 mL x4), dried and concentrated to give the crude product which was purified by preparative TLC(DCM: MeOH= 10: 1 ) to give the desired product (90 mg, 22%) as a yellow solid. LCMS: RT2.09 min; m/z 588.1 [M+H]\

H NMR (400 MHz, DMSO-d₆) δ = 8.80 - 8.71 (m, 1 H), 7.96 - 7.85 (m, 2H), 7.54-7.45 (m, 3H), 7.31 - 7.12 (m, 4H), 6.66- 6.61 (m, 1 H), 6.56 - 6.47 (m, 1 H). 5.93-5.92 (m, 1 H), 5.81- 5.75 (m, 1 H), 4.84-4.80 (m, 1 H), 4.67-4.45 (m, 2 H), 4.30-4.26 (m, 1 H), 4.25 (br s, 1 H), 3.82-3.74 (m, 5 H), 3.74 (s, 2 H), 3.68-3.50 (m, 8 H), 3.50-3.43 (m, 3 H), 3.22-3.17 (m, 2 H). 2.99(s.1 H), 2.87 (s, 1 H), 2.20-1.96 (m, 1 H).

(c) (R)^-(3-((2,3-dihydro- 1 H-inden-2-yl)(methyl)amino)-2-hydroxypropyl)-4-(morph carbonyl)benzamide 1 12

A solution of A/-(2,4-dimethoxybenzyl)-A/-((f?)-2-hydroxy-3-(4-(morpholine-4- carbonyl)benzamido) propyl)- A/-methyl-2,3-dihydro-1 H-inden-2-aminium iodide A81 (100 mg, 0.17 mmol) in trifluoroacetic acid was stirred at 70 C for 4 h. The mixture was concentrated to give the crude product. Saturated aqueous NaHC03 solution was then added and the resulting mixture was extracted with DCM (x 3). The organic layers were combined, dried and concentrated to give the crude product which was purified by preparative TLC (DCM: MeOH -10:1 ) to give the desired product (20 mg, 27%) as a yellow solid. LCMS: RT 0.46 min; m/z 438.3 [M+H]\ H NMR (400 MHz, DMSO-de) δ = 8.69 (s, 1 H), 7.95- 7.93 (m, 2H), 7.50-7.48 (m, 2H), 7.19 - 7.15(m, 4H), 4.09- 4.02 (m, 1 H), 3.63- 3.56 (m, 6H), 3.44-3.37 (m, 9 H), 3.1 1-3.13 (m, 4 H), 2.64-2.55 (m, 2 H)

Example 53: 4-(3-Oxa-9-azabicyclo[3.3.7]nonane-9-carbonyl)-/V-{(S)-3-((2,3-dihydro-1 H- inden-2- l)amino)-2-hydroxypropyl)-2-ethoxybenzamide 135

135

(a) Ethyl (3-oxa-9-azabicyclo[3.3. 1]nonane-9-carbonyl)-2-ethoxybenzoate A82

To a solution of 3-ethoxy-4-(ethoxycarbonyl)benzoic acid I70 (73 mg, 0.31 mmol, 1 equiv) in

DMF (2 mL) and acetonitrile (5 mL) was added DIPEA (160 L, 0.917 mmol, 3 equiv), HATU (174 mg, 0.458 mmol, 1 .5 equiv) and 3-oxa-9-azabicyclo[3.3. ?]nonane hydrochloride (50 mg, 0.31 mmol, 1 equiv). The reaction was stirred overnight at room temperature. The mixture was quenched with a saturated aqueous solution of NaHC03 (5 mL) and extracted with DCM (3 x 5 mL). The combined organic layers were dried in vacuo and the resultant oil was purified by column chromatography (24 g S1O2 cartridge, 0-65 % EtOAc in petroleum benzine to give the title compound (88 mg, 83% yield) as a yellow oil. LCMS-B: RT = 3.28 min, m/z = 348 [M+H]⁺.

(b) 4-(3-Oxa-9-azabicyclo[3.3.1 ]nonane-9-carbonyl) -2-ethoxybenzoic acid A83

To a solution of the ester A82 (88 mg, 0.25 mmol, 1 equiv) in THF:H₂0:MeOH (3:1 :0.5 v/v, 9 mL) was added LiOH-H₂0 (32 mg, 0.76 mmol, 3 equiv). The reaction was stirred at room temperature for 16 h before quenching by the addition of a 0.5 M aqueous solution of citric acid (10 mL) and extraction with EtOAc (3 x 15 mL). The combined organic layers were washed with water (30 mL), brine (30 mL), dried over MgS0₄ and concentrated in vacuo. The crude product mixture was purified by column chromatography (12 g S1O2 cartridge, 0- 100 % EtOAc in petroleum benzine) to give the title compound (55 mg, 68 % yield) as a colourless oil. ¹ H NMR (400 MHz, MeOD) δ 7.84 (d, J = 7.8 Hz, 1 H), 7.14 (d, J = 1.3 Hz, 1 H), 7.05 (dd, J = 7.8, 1 .4 Hz, 1 H), 4.52 (s, 1 H), 4.20 (q, J = 7.0 Hz, 2H), 4.07 - 3.99 (m, 1 H), 3.93 - 3.72 (m, 3H), 3.60 (s, 1 H), 2.02 - 1 .95 (m, obscured by EtOAc solvent), 1.92 - 1.78 (m, 2H), 1.75 - 1.64 (m, 1 H), 1.45 (t, J = 7.0 Hz, 3H), 1.40 - 1.27 (m, 2H). LCMS-B: RT = 3.00 min, m/z = 320.2 [M+H]\

(c) 4-(3-Oxa-9-azabicyclo[3.3. 1]nonane-9-carbonyl)-N-((S)-3-((2.3-dihydro- 1H-inden-2- yl)amino ) -2-hydroxypropyl) -2-ethoxybenzamide 135

To a solution of the acid A83 (83 mg, 0.26 mmol, 1 equiv), DIPEA (136 μΙ_, 0.780 mmol, 3 equiv) and HATU (148 mg, 0.390 mmol, 1 .5 equiv) in DMF (2 mL) was added a solution of the amine 11 17 (80 mg, 0.26 mmol, 1 equiv) in CH3CN (10 mL). The reaction was stirred at room temperature overnight, quenched with a saturated aqueous solution of NaHC03 (15 mL) and extracted with EtOAc (3 ^χ 40 mL). The combined organic layers were washed with water (50 mL), brine (50 mL), dried over MgS0₄ and concentrated in vacuo. DCM:TFA (8 mL, 1 :1 v/v) was added and the reaction stirred at room temperature overnight. The reaction mixture was concentrated in vacuo and then purified by solid-phase extraction (2 ^χ 1 g SCX- 2 cartridge, 3 column volumes of methanol followed by 3 column volumes of 0.2 M methanolic ammonia). The basic eluate was dried in vacuo and further purified by column chromatography (12 g S1O2 cartridge, 50-100 % EtOAc (modified by the addition of 1 % v/v of 3.5 M methanolic ammonia) in petroleum benzine followed by 0 - 20 % MeOH in EtOAc modified by the addition of 1 % v/v of 2.0 M methanolic ammonia) to give the title compound (51 mg, 39 % yield, 95 % purity by ¹H NMR) as a colourless oil. H NMR (400 MHz, MeOD) 5 8.02 (d, J = 7.9 Hz, 1 H), 7.23 - 7.15 (m, 3H), 7.13 - 7.03 (m, 3H), 4.52 (s, 1 H), 4.27 (q, J = 7.0 Hz, 2H), 4.06 - 3.76 (m, 5H), 3.68 - 3.55 (m, 3H), 3.53 - 3.43 (m, 1 H), 3.24 - 3.12 (m, 2H), 2.87 - 2.51 (m, 5H), 2.00 - 1 .95 (m, 2H), 1 .92 - 1.79 (m, 2H), 1.76 - 1.63 (m, 1 H), 1.53 (t, J = 7.0 Hz, 3H). LCMS-B: RT = 2.91 min, m/z = 508.3 [M+H]\

Example 54: 4-((1 /?,5S)-3-Oxa-9-azabicyclo[3.3.1]nonane-9-carbonyl)-A -{(S)-3-((2,3- dihydro-1 W-inden-2-yl)amino)-2-hydroxypropyl)benzamide 136

(a) Methyl (R)-4-((3-((tert-butoxycarbonyl)(2, 3-dihydro- 1 H-inden-2-yl)amino)-2- hydroxypropyl)carbamoyl)benzoate A84

To a solution of 4-(methoxycarbonyl)benzoic acid (176 mg, 0.979 mmol, 1 equiv), DIPEA (512 μΙ_, 2.94 mmol, 3 equiv) and HATU (558 mg, 1 .47 mmol, 1.5 equiv) in CH₃CN (10 mL) was added tert-butyl (f?)-(3-amino-2-hydroxypropyl)(2,3-dihydro-1 H-inden-2-yl)carbamate

11 17 (300 mg, 0.979 mmol, 1 equiv). The reaction was stirred at room temperature overnight, quenched with a saturated aqueous solution of NaHC03 (15 mL) and extracted with EtOAc (3 x 40 mL). The combined organic layers were washed with water (50 mL), brine (50 mL), dried over MgSC and concentrated in vacuo. The crude material was purified by column chromatography (12 g Si0₂ cartridge, 0-100 % EtOAc in 40 C-60 °C petroleum benzine) to give the title compound A84 (384 mg, 84 % yield) as a pale yellow foam. ¹H NMR (400 MHz, CDC ) δ 8.07 (d, J = 8.4 Hz, 2H), 7.87 (d, J = 8.4 Hz, 2H), 7.61 (s, 1 H), 7.21 - 7.08 (m, 4H), 4.64 (p, J = 8.2 Hz, 1 H), 4.00 - 3.82 (m, 5H). 3.78 - 3.71 (m, 1 H), 3.53 - 3.26 (m, 3H), 3.24 - 2.99 (m, 4H), 1 .39 (s, 9H). LCMS-B: RT = 3.43 min, m/z = 467.2 [M-H]^", 369.2 [M+2H- Boc]\

(b) (R)-4-((3-((tert-butoxycarbonyl)(2, 3-dihydro- 1 H-inden-2-yl)amino)-2- hydroxypropyl)carbamoyl)benzoic acid A85 To a solution of the ester A84 (384 mg, 0.820 mmol, 1 equiv) in THF:H₂0:MeOH (3:1 :0.5 v/v, 9 mL ) was added LiOH'h O (59 mg, 2.5 mmol, 3 equiv). The reaction was stirred at room temperature overnight before the reaction was quenched by the addition of a 0.5 M aqueous solution of citric acid (10 mL). The solution was extracted with EtOAc (3 x 15 mL). The combined organic layers were washed with water (30 mL), brine (30 mL), dried over MgSCU and concentrated in vacuo to give the title compound A85 (358 mg, 96% yield) as a white foam. LCMS-B: RT = 3.27 min, m/z = 355.1 [M+2H-Boc] ' .

(c) 4-(( 1 Ft.5S)-3-oxa-9-azabicyclo[3.3.1 ]nonane-9-carbonyl)-N-((S)-3-((2,3-dihydro- 1 H-inden- 2-yl)amino)-2-hydroxypropyl)benzamide 136

To a solution of (S)-4-((3-((2,3-dihydro-1 H-inden-2-yl)amino)-2- hyd roxypropyl )ca rba moyl )benzoic acid A85 (50 mg, 0.1 1 mmol, 1 equiv), DIPEA (57 L, 0.33 mmol, 3 equiv) and HATU (63 mg, 0.17 mmol, 1.5 equiv) in DMF (2 mL) was added a solution of the amine 3-oxa-9-azabicyclo[3.3.1]nonane hydrochloride (18 mg, 0.1 1 mmol, 1 equiv) in CH₃CN (10 mL). The reaction was stirred at room temperature overnight, quenched with a 1 .0 M solution of aqueous NaOH (15 mL) and extracted with EtOAc (3 * 40 mL). The combined organic layers were washed with water (50 mL), brine (50 mL), dried over MgSCU and concentrated in vacuo. DCM:TFA (8 mL, 1 :1 v/v) was added and the reaction stirred at room temperature for 60 h. The reaction mixture was concentrated in vacuo and then purified by solid-phase extraction (2 ^χ 1 g SCX-2 cartridge, 3 column volumes of methanol followed by 3 column volumes of 0.2 M methanolic ammonia). The basic eluate was dried in vacuo and further purified by column chromatography (12 g S1O2 cartridge, 50-100 % EtOAc (modified by the addition of 1 % v/v of 3.5 M methanolic ammonia) in petroleum benzine followed by 0 - 30 % MeOH in EtOAc modified by the addition of 1 % v/v of 3.5 M methanolic ammonia) to give the title compound (34 mg, 67 % yield) as a glassy solid. ¹ H NMR (400

MHz, MeOD) δ 7.92 (d, J = 8.3 Hz, 2H), 7.54 (d, J = 8.3 Hz, 2H), 7.22 - 7.15 (m, 2H), 7.15 - 7.06 (m, 2H), 4.54 (s, 1 H), 4.16 - 3.76 (m, 5H), 3.67 (p, J = 7.0 Hz, 1 H). 3.61 - 3.55 (m, 1 H), 3.53 - 3.45 (m, 1 H), 3.27 - 3.16 (m, 2H), 2.90 - 2.79 (m, 3H). 2.73 (dd, J = 12.2, 8.1 Hz, 1 H), 2.69 - 2.51 (m, 1 H), 2.08 - 1 .76 (m, 5H), 1 .74 - 1.62 (m, 1 H). LCMS-B: RT = 2.83 min, m/z = 464.2 [M+H]\ Purity ~ 95% by ¹H-NMR. Example 55

11 17

To a solution of the acid (0.17 mmol, 1 equiv), DIPEA (89 μΙ_, 0.51 mmol, 3 equiv) and HATU (98 mg, 0.26 mmol, 1 .5 equiv) was added a solution of ie -butyl (f?)-(3-amino-2- hydroxypropyl)(2.3-dihydro-1 H-inden-2-yl)carbamate 11 17 (52 mg, 0.17 mmol, 1 equiv) in DMF (2 mL). The reaction was stood at room temperature for 16 hours, quenched with a saturated aqueous solution of NaHCOa (5 mL) and extracted with DCM (x 3) utilizing a phase separation cartridge. The organic filtrates were reduced under a stream of air, DCM:TFA (4 mL, 1 :1 v/v) was added and the reaction stood at room temperature for the specified time. The reaction mixture was concentrated under a stream of air and then purified by solid-phase extraction (1 g SCX-2 cartridge, 3 column volumes of methanol followed by 3 column volumes of 0.2 M methanolic ammonia) to give the title compound.

Example 56: 3-((3-Acetyl-3-azabicyclo[3.2.1]octan-8-yl)amino)-W-{(S)-3-((2,3-dihydro- 1 H-inden-2- l)amino)-2-hydroxypropyl)benzamide 139

(a) Methyl 3-((3-benzyl-3-azabicyclo[3.2. 1 ]octan-8-yl)amino)benzoate A86

Methyl 3-aminobenzoate (0.100 g, 0.66 mmol) and (1 5S)-3-benzyl-3- azabicyclo[3.2.1]octan-8-one (0.214 g, 0.99 mmol) were dissolved in dry DCE (5 mL) under an atmosphere of nitrogen and acetic acid (0.076 mL, 1 .32 mmol) was added followed by sodium triacetoxyborohydride (0.280 g, 1 .32 mmol). The reaction was then stirred at room temperature for 20 hours. The reaction was quenched by addition of water (5 mL) and then diluted with EtOAc (50 mL) and saturated aqueous NaHCC>3 (50 mL). The organic phase was washed with brine (50 mL), dried (Na2SC¼), filtered and concentrated in vacuo. The crude product was purified by column chromatography (Isolera Biotage: 12 g S1O2 cartridge, 0-30% EtOAc in petroleum benzine 40-60 C) to give the title compound (0.230 g, 99% yield) as a pale yellow gum. LCMS (LCMS-B): RT 3.406 min; m/z 351 .2 [M+ H]\

(b) Lithium 3-((3-benzyl-3-azabicyclo[3.2. 1 ]octan-8-yl)amino)benzoate A87

Methyl 3-((3-benzyl-3-azabicyclo[3.2.1]octan-8-yl)amino)benzoate A86 (0.230 g, 0.656 mmol) was dissolved in THF (7 mL), MeOH (1 mL) and water (1 mL) and lithium hydroxide monohydrate (0.275 g, 6.563 mmol) was added. The reaction was then stirred at room temperature for 48 hours. The volatiles were removed in vacuo and the resulting residue diluted with EtOAc (50 mL) followed by 1 M aqueous NaOH (50 mL). The layers were separated and the organic layer was concentrated in vacuo to give the title compound (0.200 g, 89% yield) as a pale yellow gum. LC-MS (LCMS-B): RT 3.298 min; m/z 337.3 [M+H]⁺ as the free acid.

(c) 3-((3-Benzyl-3-azabicyclo[3.2. 1 ]octan-8-yl)amino)-N-((R)-3-((2, 3-dihydro- 1 H-inden-2- yl)(2,4-dimethoxybenzyl)amino)-2-hydroxypropyl)benzamide A88

Lithium 3-((3-benzyl-3-azabicyclo[3.2.1]octan-8-yl)amino)benzoate A87 (0.200 g, 0.584 mmol) was dissolved in DCM (5 mL) and DMF (1 mL) and DIPEA (0.278 mL, 1.593 mmol) followed by HATU (0.263 g, 0.690 mmol) were added. The mixture was then stirred at room temperature for 10 min before (fl)-1 -amino-3-((2, 3-dihydro- 1 H-inden-2-yl)(2,4- dimethoxybenzyl)amino)propan-2-ol 110 (0.189 g, 0.531 mmol) in DCM (2 mL) was added. The reaction was then stirred at room temperature for 24 hours. The volatiles were removed in vacuo and the resulting residue diluted with EtOAc (50 mL) followed by saturated aqueous NaHC03 (50 mL). The layers were separated and the aqueous layer was extracted with EtOAc (50 mL), the combined organic layers were washed with brine (50 mL), dried

(Na2SO.(), filtered and concentrated in vacuo to give the crude product. Purification by column chromatography (Isolera Biotage, 24 g Si Cartridge, 0-100% EtOAc in petroleum benzine 40-60 °C, then 0-20% MeOH in EtOAc) gave the title compound (0.194 g, 54% yield) as an off-white solid. LCMS (LCMS-B): RT 3.524 min, m/z 676.5 [M+H]⁺ (d) 3-((3-Acetyl-3-azabicyclo[3.2. 1 ]octan-8-yl)amino)-N-((S)-3-((2.3-dihydro- 1 H-inden-2- yl)amino)-2-hydroxypropyl)benzamide 139

3-(3-Benzyl-3-azabicyclo[3.2.1]octan-8-ylamino)-A/-(( )-3-((2,3-dihydro-1 H-inden-2-yl)(2,4- dimethoxybenzyl)amino)-2-hydroxypropyl)benzamide A88 (0.190 g, 0.282 mmol) was dissolved in EtOH (8 mL) under an atmosphere of nitrogen and 4 drops of concentrated HCI were added followed by 10% Pd/C (wet with water, 53%) (0.070 g) in EtOAc (4 mL). The atmosphere was changed to hydrogen and the flask was fitted with a hydrogen balloon (1 atm). The reaction was then stirred at room temperature for 24 hours then the atmosphere was changed to nitrogen and additional 10% Pd/C (wet with water, 53%) (0.070 g) in EtOAc (4 mL) was added. The atmosphere was again changed to hydrogen and the flask was fitted with a hydrogen balloon (1 atm). The reaction was then stirred at room temperature for 5 days. The reaction was filtered through Celite which was rinsed with EtOAc (-70 mL). A white precipitate was collected by filtration. Upon air-drying, the solid turned into a gum which was rinsed off the filter paper with methanol and the solution was concentrated in vacuo to give a brown foam. This residue was dissolved in DCM (3 mL) under an

atmosphere of nitrogen and DIPEA (0.043 mL, 0.246 mmol) followed by acetic anhydride (0.008 mL, 0.082 mmol) were added. The reaction was stirred at room temperature for 24 hours, then diluted with DCM (20 mL) and washed with saturated aqueous NaHCC>3 (20 mL), brine (50 mL), dried a2SC¾), filtered and concentrated in vacuo. To the resulting residue was added TFA (3 mL) and the reaction mixture was then heated to 70 C and stirred for 20 hours. The volatiles were removed in vacuo and the resulting residue diluted with EtOAc (50 mL) followed by 2 M aqueous NaOH (50 mL). The layers were separated and the aqueous layer was extracted with EtOAc (50 mL), the combined organic layers were washed with brine (50 mL), dried (IS^SC ), filtered and concentrated in vacuo. Purification by column chromatography (Isolera Biotage, 4 g SiC>2 Cartridge, 0-100% EtOAc in petroleum benzine 40-60 °C, then 0-66% MeOH (containing 1 % 2M N H₃ v/v) in EtOAc) gave the title compound (0.006 g, 21 % yield) as a brown gum. LC-MS (LCMS-B): RT 3.289 min, m/z 477.3 [M+H]⁺

Example 57: (S)-A -(3-((2,3-Dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)-3- (meth lsulfonamido) benzamide 140

A89

(a) (R)-N-(3-((2.3-Dihydro- 1 H-inden-2-yl)(2A-dimethoxybenzyl)amino)-2-h

(methylsulfonamido ) benzamide A89

3-(Methylsulfonamido)benzoic acid (0.109 g, 0.505 mmol) was dissolved in DCM (5 mL) and DI PEA (0.220 mL, 1 .262 mmol) followed by HATU (0.208 g, 0.547 mmol) were added. The mixture was then stirred at room temperature for 10 min before (f?)-1 -amino-3-((2,3-dihydro- 1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino)propan-2-ol 110 (0.150 g, 0.421 mmol) in DCM (2 mL) was added. The reaction was stirred at room temperature for 20 hours, then diluted with DCM (50 mL) and washed with saturated aqueous NaHC03 solution (50 mL), brine (50 mL), dried filtered and concentrated in vacuo to give the crude product. Purification by column chromatography (Isolera Biotage, 12 g S1O2 Cartridge, 0-100% EtOAc in petroleum benzine 40-60 C, then 0-35% MeOH in EtOAc) gave the title compound (0.092 g, 39% yield) as a pale yellow solid. LCMS (LCMS-A): RT 4.849 min, m/z 554.3[M+H]^*

(b) (S)-N-(3-((2,3-Dihydro-1H-inden-2-yl)amino)-2-hydroxypropyl)-3-(methylsulfo

benzamide 140

To (/7)-A/-(3-((2.3-dihydro-1 /+inden-2-yl)(2.4-dimethoxybenzyl)amino)-2-hydroxypropyl)-3- (methylsulfonamido)benzamide A89 (0.090 g, 0.163 mmol) was added TFA (3 mL) and the reaction mixture was heated to 70 °C and stirred at this temperature for 45 hours. The volatiles were partly removed in vacuo, methanol (10 mL) was added and the solution was loaded onto a SCX cartridge (2 g), the cartridge was washed with methanol (5 column volumes), then eluted with 2 M ammonia in methanol (3 column volumes). The eluate was then concentrated in vacuo to give the title compound (0.057 g, 88% yield) as a beige gum. LCMS (LCMS-B): RT 3.217 min, m/z 404.1 [M+H]⁺ Example 58: 4-(3-oxa-8-azabicyclo[3.2.1]octane-8-carbonyl)-A -((S)-3-((2,3-dihydro-1 H- inden-2- l)amino)-2-hydroxypropyl)-2-ethoxy-5-fluorobenzamide 141

(a) Ethyl 4-bromo-2-ethoxy-5-fluorobenzoate A90

4-Bromo-2.5-difluorobenzoic acid (1.00 g, 4.22 mmol) was suspended in DCM (20 mL) and cooled to 0 °C. Oxalyl chloride (0.543 mL, 6.33 mmol) and DMF (1 drop) were added and the mixture stirred at room temperature. After one hour the mixture was concentrated in vacuo, the residue dissolved in DCM (10 ml) and cooled to 0 °C. Absolute ethanol (10 mL) was added and the mixture stirred at room temperature. After 15 minutes the mixture was concentrated in vacuo and the residue taken up in absolute ethanol. Sodium metal (146 mg, 6.33 mmol) was dissolved in absolute ethanol (30 mL) and the solution added to the solution of the ester. After two hours a thick precipitate had formed. The mixture was diluted with dry THF (50 mL) and the stir bar replaced with a larger stir bar. After 19 hours the mixture was concentrated in vacuo, the residue slurried in ethyl acetate (75 mL) and filtered. The filtrate was concentrated in vacuo and purified by chromatography (12 g silica cartridge, 0-10% ethyl acetate/petroleum benzine) to give ethyl 4-bromo-2-ethoxy-5-fluorobenzoate A90 as a white solid (216 mg, 18% yield). LCMS-B: RT 3.53 min; m/z 245.0 [M-OEt]⁺ for ⁷⁹Br; ¹H NMR (400 MHz, CDCIa) δ 7.57 (d, J = 8.6 Hz, 1 H), 7.13 (d, J = 5.5 Hz, 1 H), 4.35 (q, J = 7.2 Hz, 2H), 4.07 (q, J = 7.0 Hz, 2H), 1 .46 (t, J = 7.0 Hz, 3H), 1.37 (t, J = 7.1 Hz, 3H).

(b) Ethyl 4-(3-oxa-8-azabicyclo[3.2. 1 ]octane-8-carbonyl) -2-ethoxy-5-fluorobenzoate A91 Palladium(ll) acetate (8 mg, 5 mol%), Xantphos (21 mg, 5 mol%), sodium carbonate (232 mg, 2.19 mmol), 3-oxa-8-azabicyclo[3.2.1 ]octane hydrochloride salt (163 mg, 1 .09 mmol) and ethyl 4-bromo-2-ethoxy-5-fluorobenzoate A90 (212 mg, 0.728 mmol) were stirred in toluene (2 mL) in a schlenk tube under nitrogen. The tube was flushed with carbon monoxide, and heated to 80 °C under carbon monoxide. After 18 hours the mixture was cooled to room temperature and diluted with ethyl acetate (20 mL). The mixture was filtered through Celite and the filtrate concentrated in vacuo. Chromatography (12 g silica cartridge, 0-60% ethyl acetate/hexanes) gave ethyl 4-(3-oxa-8-azabicyclo[3.2.1 ]octane-8-carbonyl)-2- ethoxy-5-fluorobenzoate A91 as a pale yellow film (41 mg, 16% yield). LCMS-B RT 3.23 min; m/z (positive ion) 352.1 [M+H]; ¹H NMR (400 MHz, CDCI3) δ 7.51 (d, J = 9.2 Hz, 1 H), 7.01 (d, J = 5.1 Hz, 1 H), 4.75 (d, J = 5.8 Hz, 1 H), 4.36 (q, J = 7.1 Hz, 2H), 4.10 (q, J = 7.0 Hz, 2H), 3.84 (d, J = 1 1.0 Hz, 1 H), 3.73 - 3.68 (m, 2H), 3.64 (d, J = 10.9 Hz. 1 H), 3.56 (dd, J = 1 1 .0, 1.6 Hz, 1 H), 2.13 - 1.90 (m, 4H), 1.45 (t, J = 7.0 Hz, 3H), 1.38 (t, J = 7.1 Hz, 3H).

(c) Lithium 4-(3-oxa-8-azabicyclo[3.2. 1]octane-8-carbonyl)-2-ethoxy-5-fluorobenzoate A92 A solution of ethyl 4-(3-oxa-8-azabicyclo[3.2.1]octane-8-carbonyl)-2-ethoxy-5-fluorobenzoate (41 mg, 0.12 mmol) in THF (1 mL) was stirred vigorously, and a solution of lithium hydroxide monohydrate (7.3 mg, 0.18 mmol) in water (0.5 mL) was added. After 4.5 hours the mixture was concentrated in vacuo, the residue dissolved in absolute ethanol and the mixture again concentrated in vacuo to give lithium 4-(3-oxa-8-azabicyclo[3.2.1 ]octane-8-carbonyl)-2- ethoxy-5-fluorobenzoate. LCMS-B: RT 2.96 min; m/z (negative ion) 322.1 [M-Li]-

(d) 4-(3-Oxa-8-azabicyclo[3.2.1 ]octane-8-carbonyl)-N-((S)-3-((2, 3-dihydro- 1 H-inden-2- yl)amino)-2-hydroxypropyl)-2-ethoxy-5-fluorobenzamide 141

Lithium 4-(3-oxa-8-azabicyclo[3.2.1]octane-8-carbonyl)-2-ethoxy-5-fluorobenzoate A92 (used directly from previous step, -0.059 mmol), triethyiamine (0.016 mL, 0.12 mmol), tert- butyl (/?)-(3-amino-2-hydroxypropyl)(2, 3-dihydro- 1 H-inden-2-yl)carbamate 11 17 (18 mg, 0.059 mmol) and HATU (34 mg, 0.18 mmol) in DMF (0.5 mL) were stood at room temperature. After 18 hours the mixture was diluted with water and DCM (1 .5 mL each). The organic phase was separated with a phase separation cartridge and treated with Dowex 50WX8 H⁺-form (200-400 mesh, 100 mg). The mixture was filtered and the filtrate diluted with 4.0 M HCI in dioxane (2 mL). After three hours the mixture was diluted with methanol (2 mL) and loaded onto a 1 g SCX cartridge. The cartridge was washed with methanol (15 mL) and eluted with 2 M ammonia in methanol (10 mL). The basic eluate was concentrated to afford the title compound. LCMS: RT 2.90 min; m/z (positive ion) 512.2 [M+H]

Example 59: ^-Methyl 3-((4-((3-((2,3-dihydro-1 H-inden-2-yS)amino)-2- hydroxypropyl)carbamoyl)pyridin-2-yl) amino)azetidine-1 -carboxylate bis(2,2,2- trifluoroacetate) 142

142

(a) (R)-Methyl 3-((4-((3-((2, 3-di hydro- 1 H-inden-2-yl)(2, 4-dimethoxybenzyl)amino) -2- hydroxypropyl)carbamoyl)pyridin-2-yl)amino)azetidine- 1 -carboxylate A93

To a solution of (f?)-2-(azetidin-3-ylamino)-W-(3-((2,3-dihydro-1 H-inden-2-yl)(2,4- dimethoxybenzyl)amino)-2-hydroxypropyl)isonicotinamide dihydrochloride A74 (100 mg. 0.18 mmol) in DCM (4 mL) was added triethvlamine (95 mg, 0.94 mmol). A solution of methyl chloroformate (22 mg, 0.22 mmol) in DCM (1 mL) was added dropwise into the mixture. The resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated under vacuum and the residue partitioned between DCM (10 mL) and water (10mL). The aqueous layer was extracted with DCM (3 x 5mL) and the combined organic extracts washed with brine (3 x 5mL), dried (NazSC ), filtered and concentrated. The residue was purified by preparative TLC (5% methanol in dichloromethane) to give the title compound as a white solid (53 mg, 48%). LCMS RT 2.07min; m/z 590.3 [M+H]' (b) (S)-Methyl 3-((4-((3-((2,3-dihydro-1H-inden-2-yl)amino)-2- hydroxypropyl)carbamoyl)pyridin-2-yl) amino)azetidine- 1 -carboxylate bis(2,2,2- trifluoroacetate) 142

A solution of (fl)-methyl 3-((4-((3-((2,3-dihydro-1 H-inden-2-yl)(2,4-dimethoxybenzyl)amino)- 2-hydroxypropyl)carbamoyl)pyridin-2-yl)amino)azetidine-1 -carboxylate A93 (50 mg, 0.08 mmol) in trifluoroacetic acid (4 mL) was heated at reflux 6 h. The mixture was concentrated under vacuum and the residue obtained dissolved in DCM (10 mL). The reaction was quenched with saturated aqueous NaHCC>3 solution (15 mL). The aqueous layer was extracted with DCM (3 x 10 mL) and the combined organic extracts washed with saturated aqueous NaHCOa solution (3 x 5 mL), brine (3 x 5 mL), dried (Na2SC>4), filtered and concentrated. The residue was purified by preparative HPLC to give the title compound as a yellow solid (15 mg, 40%). ¹H NMR (400MHz, MeOH-d₄) δ 8.04 (d, J = 6.4 Hz, 1 H), 7.33 (s, 1 H), 7.27-7.19 (m, 5H), 4.66-4.60 (m, 1 H), 4.45-4.40 (m, 2H), 4.14-4.08 (m, 2H), 3.98-3.95 (m, 2H). 3.68 (s, 3H), 3.57-3.53 (m, 1 H), 3.49-3.38 (m, 3H). 3.29-3.25 (m, 1 H), 3.19-3.1 1 (m, 2H). 3.07-3.02 (m, 1 H). LCMS RT 2.74 min; m/z 440.2 [M+H] ' for the free base.

Example 60: fSj-2-(azetidin-3-yIamsno)-N-(3-((2,3-dihydro-1 H-inden-2-yl)amino)-2- hydroxypropyl isonicotinamide tris(2,2,2-trifluoroacetate) 143

A solution of (/?)-2-(azetidin-3-yiamino)-A -(3-((2,3-dihydro-1 H-inden-2-yi)(2.4- dimethoxybenzyl)amino)-2-hydroxypropyl)isonicotinamide dihydrochloride A74 (70 mg, 0.13 mmol) in trifluoroacetic acid (3 mL) was heated at reflux 4 h . The mixture was concentrated under vacuum and the residue obtained dissolved in DCM (10 mL). The mixture was quenched with a saturated aqueous NaHCOa solution (10 mL). The aqueous layer was extracted with DCM (3 x 5mL) and concentrated under vacuum. The residue was dissolved in a mixed solution of methanol/dichloromethane (20 mL, 1/20 v/v) and the mixture filtered. The filtrate was concentrated and the residue purified by preparative HPLC to give the title compound as an off-white solid (16 mg, 32%): Ή NMR (400 MHz, MeOD) δ 8.25 (d, J = 6.8 Hz, 1 H), 7.49 (s, 1 H), 7.33 (d, J = 6.4 Hz, 1 H), 7.27-7.20 (m, 4H), 5.02-4.96 (m, 1 H), 4.79- 4.74 (m, 1 H), 4.68-4.63 (m, 1 H), 4.15-4.08 (m, 2H), 3.57-3.53 (m, 1 H), 3.49-3.35 (m, 5H), 3.29-3.25 (m, 1 H), 3.19-3.1 1 (m, 2H), 3.07-3.02 (m, 1 H); LCMS RT 0.27 min; m/z 382.2 [M+H]⁺ for the free base. Example 61 : 4-(3-Oxa-8-azabicyclo[3.2.1]octane-8-carbonyl)-N-((f?)-3-((2,3-dihyd inden-2-yl)amino)-2- hydroxypropyl)-2-ethoxybenzamide hydrochloride 144

(a) tert-Butyl((2R)-3-(4-(3-oxa-8-azabicyclo[3.2.1 ]octane-8-carbonyl)-2-ethoxybenzamido)-2- hydroxypropyl)(2, 3-dihydro- 1 H-inden-2-yl)carbamate A94

A mixture of (S)-iert-butyl (3-amino-2-hydroxypropyl)(2,3-dihydro-1 /-/-inden-2-yl)carbamate (1 .0 g, 3.26 mmol) 1123, 4-(3-oxa-8-azabicyclo[3.2.1]octane-8-carbonyl)-2-ethoxybenzoic acid I72 (1 .14 g, 3.92 mmol), HOBt (0.66 g, 4.89 mmol), EDCI (0.94 g, 4.89 mmol) and triethylamine (1 .32 g, 13.04 mmol) in CH2CI2 (20 mL) under 2 was stirred at room temperature overnight. The mixture was poured into water and diluted with EtOAc, the organic layer was washed with a 0.1 M aqueous HCI solution, brine, dried (Na2SO.i), filtered, concentrated and purified by silica gel chromatography (MeOH/Ch C = 1/1 , v/v) to give the title compound (1.3 g, 67%) as a white solid. LCMS: RT 3.02 min; m/z 594.4 [M+H]\ ¹H

NMR (400 MHz, CDCI3) δ 8.54 (br s, 1 H), 8.19 (d, J = 0.8 Hz, 1 H), 7.16-7.07 (m, 6H), 4.72- 4.68 (m, 2H), 4.23 (q, J = 6.8 Hz, 2H), 3.93-3.85 (m, 3H), 3.74-3.59 (m, 4H), 3.39-3.32 (m, 3H), 3.18-3.12 (m, 2H), 3.08-2.99 (m, 2H), 2.05-1.93 (m, 4H), 1.54 (t, J = 7.2 Hz, 3H), 1.41 (s. 9H).

(b) 4-(3-Oxa-8-azabicyclo[3.2.1 ]octane-8-carbonyl)-N-((R)-3-((2,3-dihydro- 1 H-inden-2- yl)amino)-2- hydroxypropyl)-2-ethoxybenzamide hydrochloride 144

To a solution of ie t-butyl ((2S)-3-(4-(3-oxa-8-azabicyclo[3.2.1 ]octane-8-carbonyl)-2- ethoxybenzamido)-2-hydroxypropyl)(2.3-dihydro-1 /+inden-2-yl)carbamate (500 mg, 0.84 mmol) in CH2CI2 (5 mL) was added dropwise a solution of HCI (2.5 M, in dioxane, 5 mL) at 0 °C. The mixture was stirred at room temperature overnight. The solvents were removed in vacuo and Et^O was added, the precipitate was collected by filtration and washed with Et^O and dried to give the title compound (435 mg, 97%) as a white solid. LCMS:RT 1.95 min, m/z 494.1 [M+H]⁺; H N MR (400 MHz, DMSO-de) δ 9.13 (br s,2H), 8.42 (t, J = 5.6Hz, 1 H), 7.83 (d, J = 8.0 Hz, 1 H), 7.26-7.1 1 (m ,5H), 5.82 (d, J = 4.2 Hz, 1 H), 4.52 (br s, 1 H), 4.23 (q, J = 6.8 Hz, 2H), 4.05-4.02 (m, 2H), 3.86 (br s, 1 H), 3.67-3.61 (m, 3H), 3.51 -3.42 (m, 3H), 3.29- 3.25 (m, 2H), 3.17-3.10 (m, 3H), 2.95-2.90 (m, 1 H), 1 .88 (br s, 4H), 1 .42 (t, J = 6.8 Hz, 3H).

Example 62: 6-(8-Oxa-3-azabicyclo[3.2.1]octan-3-yl)-/V-((S)-3-((2,3-dihydro-1 /+inden-2- l)amino)-2-hydroxypropyl)nicotinamide 145

(a) Methyl 6-(8-oxa-3-azabicyclo[3.2.1 ]octan-3-yl)nicotinate A95

To a solution of methyl 6-chloronicotinate (300 mg, 1 .75 mmol) in dry DMF (15 mL) was added 8-oxa-3-azabicyclo[3.2.1 ]octane hydrochloride (392 mg, 2.62 mmol, 1.5 equiv) and triethylamine (0.609 mL, 4.37 mmol, 2.5 equiv). The system was flushed with N2 and heated at 80 C for 20 h. The reaction was allowed to reach room temperature and was diluted with water/EtOAc. The aqueous layer was extracted with ethyl acetate (* 3). The combined organic layers were washed with brine, dried over MgS0₄, filtered, and the solvent removed in vacuo. The crude residue was purified by column chromatography (24 g S1O2 cartridge, 0- 35 % EtOAc in petroleum benzine 40 C-60 C) to give the title compound (400 mg, 92 % yield) as an off-white solid. Ή NMR (400 MHz, CDC ) δ 8.76 (dd, J = 2.3, 0.8 Hz, 1 H), 8.00 (dd, J = 9.0, 2.4 Hz, 1 H). 6.48 (dd, J = 9.1. 0.8 Hz, 1 H), 4.52 - 4.41 (m, 2H), 3.96 - 3.87 (m, 2H), 3.84 (s, 3H), 3.18 (dd, J = 12.6, 2.7 Hz, 2H), 1.99 - 1.89 (m, 2H), 1 .83 - 1.74 (m, 2H). LCMS-B: RT 3.05 min, m/z = 249.1 [M+H]\

(b) 6-(8-Oxa-3-azabicyclo[3.2.1 ]octan-3-yl)nicotinic acid A96

To a solution of the desired ester methyl 6-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)nicotinate A95 (400 mg, 1.61 mmol) in THF:H₂0:MeOH (3:1 :0.5 v/v. 10 mL) was added ϋΟΗΉ₂0 (203 mg, 4.83 mmol, 3 equiv). The reaction was stirred at room temperature for 16 h. The reaction mixture was concentrated in vacuo. Water was added and the pH adjusted to 3 by the addition of a 0.5 M aqueous citric acid solution. The aqueous mixture was extracted with ethyl acetate (x 3), washed with water, brine, dried ( a2S0₄) and the solvent removed in vacuo to give the title compound (347 mg, 92 % yield) as a light brown solid. ¹H NMR (400 MHz, DMSO) δ 8.59 (d, J = 2.3 Hz, 1 H), 7.90 (dd, J = 9.0, 2.4 Hz, 1 H), 6.73 (d, J = 9.0 Hz, 1 H), 4.41 (dd, J = 4.2, 2.1 Hz, 2H), 3.94 (d, J = 12.5 Hz, 2H), 3.01 (dd, J = 12.7, 2.6 Hz, 2H), 1.87 - 1.72 (m, 2H), 1.65 (q, J = 7.1 , 6.5 Hz, 2H). LCMS-A: RT 1.85 min, m/z = 235.2

[M+H]\

(c) 6-(8-Oxa-3-azabicyclo[3.2. 1]octan-3-yl)-N-((S)-3-((2,3-dihydro^

hydroxypropyl)nicotinamide 145

To a solution of ieri- butyl (f?)-(3-amino-2-hydroxypropyl)(2, 3-d ihydro-1 H-inden-2- yl )carbamate 11 1 7 (100 mg, 0.326 mmol), DIPEA (171 μΙ_. 0.979 mmol, 3 equiv) and HATU (186 mg, 0.490 mmol, 1 .5 equiv) in DMF (2 mL) was added a solution of the 6-(8- oxa-3-azabicyclo[3.2.1 ]octan-3-yl)nicotinic acid A96 (76 mg, 0.33 mmol, 1 equiv) in DMF (3 mL). The reaction was stirred at room temperature overnight, quenched with a 1 M aqueous solution of NaOH (15 mL) and stirred for 4 h. The aqueous mixture was extracted with EtOAc (3 x 40 mL) and the combined organic layers were washed with water (50 mL), brine (50 mL), dried over MgS0₄ and concentrated in vacuo. DCM:TFA (8 mL, 1 :1 v/v) was added and the reaction stirred at room temperature overnight. The reaction mixture was concentrated in vacuo and then purified by solid-phase extraction (5 g SCX-2 cartridge, 3 column volumes of methanol followed by 3 column volumes of 0.2 M methanolic ammonia). The basic eluate was dried in vacuo and further purified by column

chromatography (12 g Si0₂ cartridge, 50-100 % EtOAc (modified by the addition of 1 % v/v of 3.5 M methanolic ammonia) in petroleum benzine followed by 0 - 20 % MeOH in EtOAc modified by the addition of 1 % v/v of 3.5 M methanolic ammonia) to give the title compound 6-(8-oxa-3-azabicyclo[3.2.1 ]octan-3-yl)-A/-((S)-3-((2,3-dihydro-1 H-inden-2- yl)amino)-2-hydroxypropyl)nicotinamide (56 mg, 41 % yield) as a white solid. ¹H NMR (400 MHz, MeOD) δ 8.60 (dd, J = 2.5, 0.7 Hz, 1 H). 7.95 (dd, J - 9.0, 2.5 Hz, 1 H), 7.25 - 7.16 (m, 2H), 7.16 - 7.06 (m, 2H), 6.74 (dd, J = 9.2, 0.8 Hz, 1 H), 4.51 - 4.42 (m, 2H), 4.01 - 3.90 (m, 3H), 3.73 (p. J = 7.0 Hz, 1 H), 3.51 - 3.38 (m, 2H), 3.28 - 3.18 (m, 2H), 3.12 (dd, J = 12.7, 2.6 Hz, 2H), 2.94 - 2.82 (m, 3H), 2.76 (dd, J = 12.3, 8.2 Hz, 1 H), 2.00 - 1.91 (m, 2H), 1.86 - 1.73 (m, 2H). LCMS-B: RT 2.83 min, m/z = 423.2 [M+H]\ Example 63: (S)-/V-(3-((2,3-Dihydro-1 H-inden-2-yl)amino)-2-hydroxypropyl)-2-ethoxy-6- morpholinonicotinamide 146

(a) 2-Ethoxy-6-morpholinonicotinic acid A97

6-Chloro-2-ethoxynicotinic acid (0.250 g, 1.24 mmoi) was dissolved in 1.4-dioxane (2 mL) and ΕΪ3Ν (0.52 mL, 3.72 mmol) and morpholine (0.16 mL, 1 .86 mmol) were added and the reaction was heated in a microwave at 150 C for 45 minutes. The solvent was evaporated and the residue was purified by chromatography (Isolera Biotage, 12 g Si02 cartridge, 0-100 EtOAc in petroleum benzine 40-60 °C, then 0-5% MeOH in EtOAc) to give the title compound (0.174 g, 56% yield) as a pale pink solid. ¹ H NMR (400 MHz, MeOD) δ 8.05 (d, J = 8.7 Hz, 1 H), 6.36 (d, J = 8.7 Hz, 1 H), 4.47 (q, J = 7.1 Hz, 2H), 3.79 - 3.73 (m, 4H), 3.67 - 3.62 (m, 4H), 1.40 (t, J = 7.0 Hz, 3H). LCMS-B: RT 3.098 min, m/z 253.1 [M+H] ^* (b) tert-Butyl (R)-(2, 3-dihydro- 1 H-inden-2-yl)(3-(2-ethoxy-6-morpholinonicotinamido)-2- hydroxypropyl)carbamate A98

2-Ethoxy-6-morpholinonicotinic acid A97 (0.050 g, 0.198 mmol) was dissolved in DMF (4 mL) and Et₃N (0.083 mL, 0.595 mmol) followed by HATU (0.1 13 g, 0.297 mmol) were added. The mixture was then stirred at room temperature for 10 minutes before iert-butyl (R)-(3- amino-2-hydroxypropyl)(2,3-dihydro-1 /-/-inden-2-yl Carbamate 11 17 (0.061 g, 0.198 mmol) was added. The reaction was stirred at room temperature for 20 hours, then diluted with EtOAc (70 mL) and washed with saturated aqueous NaHC03 solution (70 mL), brine (70 mL), dried ( a2S0₄), filtered and concentrated in vacuo to give the crude product. This was purified by column chromatography (Isolera Biotage, 12 g S1O2 cartridge, 0-80% EtOAc in petroleum benzine 40-60 °C) to give the title compound (0.073 g, 68% yield) as a colourless gum. LCMS-B: RT 3.552 min, m/z 541.3 [M+H] ' (c) (S)-N-(3-((2, 3-Dihydro- 1 H-inden-2-yl)amino)-2-hydroxypropyl)-2-ethoxy-6- morpholinonicotinamide 146

iert-Butyl (f?)-(2,3-dihydro-1 H-inden-2-yl)(3-(2-eihoxy-6-morpholinonicoiinamido)-2- hyd roxypropyi )ca rba mate A98 (0.073 g, 0.135 mmol) was dissolved in DCM (4 mL), TFA (0.310 mL, 4.051 mmol) was added and the reaction was stirred at room temperature for 20 hours. Volatiles were removed in vacuo and the residue was diluted with EtOAc (70 mL) and washed with 2 M aqueous NaOH solution (70 mL), brine (70 mL), dried (Na2S04), filtered and concentrated in vacuo, taken up in diethyl ether and evaporated in vacuo to give the title compound (0.053 g, 89% yield) as a white foam. ¹H NMR (400 MHz, MeOD) δ 8.14 (d, 1 H), 7.20 - 7.08 (m, 4H), 6.41 (d, J = 8.7 Hz, 1 H), 4.54 - 4.47 (m, 2H), 3.95 - 3.87 (m, 1 H), 3.79 - 3.74 (m, 4H), 3.69 - 3.54 (m, 6H), 3.42 (dd, J = 13.8, 6.4 Hz. 1 H), 3.19 (ddd, J = 15.7, 7.2, 5.3 Hz, 2H), 2.85 - 2.76 (m, 3H), 2.71 (dd, J = 12.1 , 8.0 Hz, 1 H), 1.46 (t, J = 7.1 Hz, 3H). LCMS-B: RT 2.981 min, m/z 441 .2 [M+H]⁺ Assays

PRMT5 Biochemical Assay

Compounds of the invention may be tested for in vitro activity in the following assay:

A histone H4 derived peptide is used as substrate (amino acid sequence: Ser-Gly-Arg-Gly- Lys-Gly-Gly-Lys-Gly-Leu-Gly-Lys-Gly-Gly-Ala-Lys-Arg-His-Arg-Lys-Val-NH₂). Full-length PRMT5 enzyme (NCBI Reference sequence NP_006100.2) was co-expressed with Hise- MEP50 in insect cells and purified via Nickel immobilized metal affinity and gel filtration chromatography ("the enzyme").

The 6 L assay reactions are run in Greiner brand black 384-well low volume plates. All reactions contained assay buffer (phosphate buffered saline, 0.01 % (v/v) Tween-20, 0.01 % (w/v) albumin from chicken egg white, 1 mM dithiothreitol, 200 nM peptide substrate, 1 μΜ S-Adenosyl methionine, and 15 ng/reaction enzyme, with the enzyme being omitted from negative control reactions. Compounds were added in a volume of 100 nL from dilution series prepared in DMSO, positive and negative control reactions receiving the same volume DMSO without compound. The plates were sealed with adhesive seals and incubated for 4 hours at 37 °C. Reaction progress was measured using the Transcreener™ EPIGEN methyltransferase assay (BellBrook Labs, Madison, Wl) as recommended by the

manufacturer. To each reaction 2 L detection mix were added, containing coupling enzymes, fluorescence polarisation tracer, and AMP antibody. Plates were incubated for 90 min before being read on a PerkinElmer En Vision™ plate reader in fluorescence polarisation mode. ICso values were obtained from the raw readings by calculating percent inhibition (%l) for each reaction relative to controls on the same plate (%I=(I-CN)/(CP-CN) where CN/ CP are the averages of the negative/ positive reactions, respectively), then fitting the %l data vs. compound concentration [I] to %I=(A+((B-A)/(1 +((C/[I])^AD)))) where A is the lower asymptote, B is the upper asymptote, C is the IC50 value, and D is the slope.

Compound Number IC50 (μ )

1 0.792

2 0.31

3 0.309

4 3.79

5 9.78

6 1.55

7 2.45

8 0.263

9 3.72

10 1.44

1 1 0.83

12 0.695

13 0.634

14 0.445

15 0.218

16 0.301

17 0.629

18 0.238

20 0.977

21 1.1 1 1

22 0.171

24 0.425

25 2.22

26 3.87

27 0.434

28 69.4

29 4.43

30 0.905

31 22.58 Compound Number ICso (μ )

32 18.8

33 24.9

34 41 .1

35 0.050

37 0.095

38 0.210

39 0.120

40 0.057

41 0.157

PR T5 Biomarker Assay

Compounds of the invention may be tested for potency to inhibit symmetrical dimethylation of arginine in the following assay:

The cell line TE1 1 was seeded at a density of 12.000 cells per well in 96 well tissue culture plates in DME medium and 10% foetal bovine serum, and allowed to adhere overnight under standard culture conditions (37 C, 5 % CO2). Compound dilutions prepared in DMSO were added to the medium, with negative control wells reserved for treatment with DMSO only and positive controls receiving a potent PRMT5 inhibitor. The concentration of the inhibitor had been previously determined to give maximum inhibition of the methylation. After incubation for 72 h, cells were washed twice in ice-cold PBS, lysed in lysis buffer (20 mM Tris pH 7.4, 135 mM NaCI, 1 .5 mM MgC . 1 mM EGTA, 10 % glycerol and 1 % Triton-X100), centrifuged at 15,000 xg and the supernatants collected for subsequent analysis. The methylation level was determined using the EpiQuik™ Global Di-Methyl Histone H4R3 Quantification ELISA Kit (Epigentek, Farmingdale, NY) as per the manufacturer's

recommendations; in parallel the total protein amount in the lysate was quantified using a Lowry protein assay. The methylation level was corrected for the total protein amount of each sample, normalised to the controls, and the data fitted against a four-parameter logistic model to determine the 50% inhibitory concentration (IC50).

Compound Number ICso (μΜ)

2 0.069

8 0.336

13 0.348

14 0.21 15 0.030

16 0.259

18 0.092

22 0.018

24 0.363

30 1.910

35 0.008

37 0.019

38 0.01 1

40 0.002

98 0.007

Revised PRMT5 Biochemical Assay

Compounds of the invention may be tested for in vitro activity in the following assay:

A histone H4 derived peptide is used as substrate (amino acid sequence: Ser-Gly-Arg-Gly- Lys-Gly-Gly-Lys-Gly-Leu-Gly-Lys-Gly-Gly-Ala-Lys-Arg-His-Arg-Lys-Val-NH₂). Full-length PRMT5 enzyme (NCBI Reference sequence P_006100.2) was co-expressed with Hise- MEP50 in insect cells and purified via Nickel immobilized metal affinity and gel filtration chromatography ("the enzyme"). The 6 μΙ_ reactions are run in Greiner brand black 384-well low volume assay plates. All reactions contained assay buffer (phosphate buffered saline, 0.01 % (v/v) Tween-20, 0.01 % (w/v) albumin from chicken egg white, 1 mM Dithiothreitol, 1 μΜ peptide substrate, 1 μΜ S-Adenosyl methionine, and 15 ng/reaction enzyme, with the enzyme being omitted from negative control reactions. Compounds were added in a volume of 100 nl_ from dilution series prepared in DMSO, positive and negative control reactions receiving the same volume DMSO without compound. The plates were sealed with adhesive seals and incubated for 4 hours at 37 degree Celsius. Reaction progress was measured using the Transcreener™ EPIGEN methyltransferase assay (BellBrook Labs, Madison, Wl) as recommended by the manufacturer. To each reaction 2 μΙ_ detection mix were added, containing coupling enzymes, fluorescence polarisation tracer, and AMP antibody. Plates were incubated for 90 minutes before being read on a PerkinElmer En Vision™ plate reader in fluorescence polarisation mode. IC50 values were obtained from the raw readings by calculating percent inhibition (%l) for each reaction relative to controls on the same plate (%I=(I-CN)/(CP-CN) where CN/ CP are the averages of the negative/ positive reactions, respectively), then fitting the %l data vs. compound concentration [I] to %I=(A+((B-A)/(1 +((C/[I])^AD)))) where A is the lower asymptote, B is the upper asymptote, C is the IC50 value, and D is the slope.

Compound Number IC50 (μΜ)

35 0.155

37 0.163

38 0.310

42 0.137

43 1.010

44 0.373

45 12.822

46 4.988

47 1 1 .441

48 4.784

49 25.309

50 0.466

51 0.709

52 74.637

53 0.164

54 5.769

55 0.450

56 0.098

57 2.533

58 0.883

59 0.487

60 0.630

61 0.180

62 6.925

63 7.295

64 1.104

65 1.299

66 1.559

67 6.871

68 6.171

69 10.004 Compound Number ICso (μΜ)

70 0.157

71 0.237

72 0.341

73 4.379

74 3.135

75 0.064

76 0.209

77 0.029

78 5.577

79 24.001

80 0.253

81 1.366

82 1.813

83 3.458

84 0.585

85 7.684

86 0.682

87 2.762

88 0.229

89 0.853

90 3.153

91 1.226

92 1.383

93 0.505

94 1.346

95 0.931

96 0.733

97 4.319

98 0.281

100 1.104

101 0.457

102 0.482

103 0.120

104 0.514 Compound Number ICso (μΜ)

105 0.196

106 24.254

107 52.161

108 21.239

109 11.105

110 0.166

111 10.382

112 9.413

113 4.146

114 0.066

115 0.305

116 0.276

117 31.368

118 102.627

119 5.082

120 13.317

121 0.130

122 0.166

123 0.040

124 0.048

125 6.724

126 0.226

127 0.137

128 15.438

129 0.629

130 1.129

131 4.591

132 29.683

133 0.131

134 0.238

135 0.015

136 0.050

137 46.992

138 16.890 Compound Number ICso (μΜ)

139 0.771

140 4.679

141 0.695

Revised PRMT5 Biomarker Assay

Compounds of the invention may be tested for potency to inhibit symmetrical dimethylation of arginine in the following assay:

The cell line TE1 1 was seeded at a density of 6,000 cells per well in 96 well optical quality tissue culture plates in DME medium and 10% foetal bovine serum, and allowed to adhere for 5 hours under standard culture conditions (37 degree Celsius, 5% CO2). Compound dilutions prepared in DMSO were added to the medium, with negative control wells reserved for treatment with DMSO only and positive controls receiving a potent PRMT5 inhibitor compound at 1 μΜ concentration. After incubation for 72 hours, the cells were fixed with 3.7% formaldehyde in PBS for 30 minutes at room temperature, washed with phosphate buffer saline and blocked with Odyssey blocking buffer (LI-COR, Lincoln, NE). Rabbit anti- Di-Methyl Histone H4 Arginine 3 specific antibody (Epigentek) in Odyssey blocking buffer was added and incubated for 14 hours at 4 degree Celsius. After washing, anti-rabbit secondary antibody labelled with Alexa647 dye (LifeTechnologies) and Hoechst 33342 (1 pg/mL, SigmaAldrich) were added for 1 hour incubation. Plates were washed and read on a PerkinElmer Envision 2103 in fluorescence intensity scanning mode (24 scans across the well area). The methylation level information was corrected for the number of cells as expressed by the Hoechst 33342 stain, converted to percent inhibition relative to controls on the same plate and the data fitted against a four-parameter logistic model to determine the 50% inhibitory concentration (IC50 plate-reader based). Alternatively, the plates were imaged on a PerkinElmer Phenix high content imaging platform. Using a Columbus image analysis pipeline, individual nuclei were located by Hoechst 33342 stain and the methylation level was calculated from the Alexa647-related intensity in the same area. The resulting mean intensity per cell was directly converted to percent inhibition as outlined above (IC50, imager based)). Compound Number ICso (μΜ) - plate reader based

15 0.025

22 0.106

35 0.040

38 0.053

40 0.021

53 0.662

55 0.122

56 0.007

70 0.015

75 0.004

76 0.008

77 0.0005

98 0.361

103 0.017

105 0.010

1 1 1 >2

1 14 0.024

121 0.016

123 0.012

124 0.013

127 0.002

133 0.009

135 0.0003

Claims

wherein:

n is 1 or 2;

R^N is H or Me;

R¹ is optionally one or more halo or methyl groups;

R^2a and R^2b are independently selected from the group consisting of:

(i) F;

(ii) H;

(iii) Me; and

R^2c and R^2d (if present) are independently selected from the group consisting of:

(i) F;

(ii) H;

(iii) Me; and

(iv) CH₂OH;

R^3a and R^3b are independently selected from H and Me;

R^4a is selected from OH, -NH₂, -C(=0)NH₂, and -CH2OH;

R^4b is either H or Me;

R⁵ is either H or Me; A is either

(i) optionally substituted phenyl;

(ii) optionally substituted naphthyl; or

(iii) optionally substituted C5-12 heteroaryl.

2. A compound according to claim 1 , wherein n is 1.

3. A compound according to claim 1 , wherein n is 2.

4. A compound according to any one of claims 1 to 3, wherein R^N is H.

5. A compound according to any one of claims 1 to 3, wherein R^N is Me.

6. A compound according to any one of claims 1 to 5, wherein there are no R¹ substituents.

7. A compound according to any one of claims 1 to 5, wherein R¹ represents one to four Me or halo groups.

8. A compound according to any one of claims 1 to 7, wherein R^2a, R^2b, R^2c and R^2d (if present) are all H.

9. A compound according to any one of claims 1 to 7 of formula la or lb, wherein R^2a, R^2b, R^2c and R^2d are comprised of three H and one Me or CH2OH group.

10. A compound according to claim 9 of formula la or lb, wherein R^2a is Me and R^2b, R^2c and R^2d are H.

1 1 . A compound according to claim 9, wherein R^2c is Me or CH2OH and R^2a, R^2b and R^2d are H.

12. A compound according to any one of claims 1 to 7 of formula la or lb, wherein R ^a, R² , R ^c and R^2d are comprised of two H and two Me groups.

13. A compound according to claim 12, wherein R^2a and R^2c are Me and R^2b and R^2d are H.

14. A compound according to claim 12, wherein R^2a and R^2b are Me and R^2c and R^2d are H.

15. A compound according to claim 12, wherein R^2c and R^2d are Me and R ^a and R^2b are H.

16. A compound according to any one of claims 1 to 7 of formula Ic, wherein R^2a is H and R^2b is a Me or CH₂OH group.

17. A compound according to any one of claims 1 to 7 of formula Ic, wherein R^2a and R^2b are each a Me or CH2OH group.

18. A compound according to any one of claims 1 to 17, wherein R^3a is H and R^3b is Me.

19. A compound according to any one of claims 1 to 17, wherein R^3a and R^3b are both H.

20. A compound according to any one of claims 1 to 17, wherein R^3a and R^3b are both Me.

21 . A compound according to any one of claims 1 to 20, wherein R^4a is OH.

22. A compound according to any one of claims 1 to 21 , wherein R^4b is H.

23. A compound according to any one of claims 1 to 21 , wherein R'^, is Me.

24. A compound according to any one of claims 1 to 23, wherein R⁵ is H.

25. A compound according to any one of claims 1 to 23, wherein R⁵ is Me.

26. A compound according to claim 1 which is of formula Ia1 , Ib1 or Id :

A compound accordin to claim 1 which is of formula Ia2, Ib2 or Ic2

28. A compound according to any one of claims 1 to 27, wherein the compound is a racemate at the carbon atom to which R^4b and R^4a are attached.

29. A compound according to any one of claims 1 to 29, wherein the compound is a single enantiomer at the carbon atom to which R^4b and R^4a are attached.

30. A compound according to claim 29, wherein the compound is the ff -enantiomer.

31 . A compound according to claim 29, wherein the compound is the fSj-enantiomer.

32. A compound according to any one of claims 1 to 31 , wherein the optional

substituents on A are independently selected from the group consisting of: C alkyl, CM fluoroalkyi, C3-6 cycloalkyi, C5-6 heteroaryl, C5-6 heteroaryl methyl, C4-6 heterocyclyl, C4-6 heterocyclyl methyl, phenyl, benzyl, halo, amido, amidomethyl, acylamido, acylamidomethyl, C1-4 alkyl ester, C1-4 alkyl ester methyl, CM alkyl carbamoyl, CM alkyl carbamoyl methyl, CM alkylacyl, CM alkylacyl methyl, phenylcarbonyl, carboxy, carboxymethyl, ether, amino, aminomethyl, sulfonamido, suifonamino, suifone, sulfoxide, nitrile and nitriiemethyl and when A is phenyl, the optional substituent may also be a fused C5-6 Nrcontaining heterocyclic ring.

33. A compound according to any one of claims 1 to 32, wherein A is optionally substituted phenyl.

34. A compound according to claim 33, wherein A is unsubstituted phenyl.

35. A compound according to claim 33, wherein A is phenyl having 1 , 2. 3. 4 or 5 substituents.

36. A compound according to claim 35, wherein the phenyl has 1 or 2 substituents.

37. A compound according to claim 35, wherein the phenyl has 1 substituent.

38. A compound according to claim 35, wherein the phenyl has 2 substituents.

39. A compound according to any one of claims 35 to 38, wherein the substituents are selected from: CM alkyl, fluoro, chloro, bromo, acetyl, methoxy, ethoxy, -C(=0)Me, - C(=0)Et, -CH₂C(=0)Me, phenyl, -CF₃, -CF₂H, -CN, -CH₂CN, -OBn. -OPh, -OCF₃,-OCF₂H, - 0-(C₆H._t)-CN, -COOH, -CH2COOH, -C(=0)OMe, -C(=0)NH₂, -C(=0)NMeH, -C(=0)NMe₂, - C(=0)N'PrH, -C(=0)-piperidinyl, -C(=0)-pyrrolidinyl.-C(=0)-morpholino (which may be bridged or substituted with one or two methyl groups), -C(=0)-azetidinyl, -CH₂C(=0)NH₂, - CH₂C(=0)-azetidinyl, -CH₂C(=0)NMeH, -CH₂C(=0)N'PrH, -CH₂C(=0)-pyrrolidinyl, - CH₂C(=0)-morpholino, -CH₂-morpholino, -CH₂-methylpiperazinyl, -OCH₂pyridinyl, - OCH₂-methyloxadiazolyl, -CH₂-imidazolyl, -O-tetrahydropyranyl, -CH₂-tetraydropyanyl.-NH- methylpyrazinyl, -CH₂-triazolyl, -NHS0₂Ph, -NHS0₂Me, -S0₂NMePh, -S0₂NMe , -S0₂NHEt, -SO2CF3,- γ-lactam, -CH₂NHC(=0)Me, -CH₂NHC(=0)OMe, -CH₂NHC(=0)CF₃, morpholino, - CH2NH2, -C(=0)Ph, -OCH2-isoxazolyl, -NH-pyrimidinyl, pyridizinyl, pyrimidinyl, pyridinyl, pyrazolyl, methylpyrazolyl, dimethylpyrazolyl, pyrazinyl, pyridazinyl , methyloxadiazolyl, oxadiazoiyi, dimethyloxadiazoiyi, isoxazolyl, dimethyitriazoiyi, imidazoiyi, benzimidazoiyi and thiadiazolyl.

40. A compound according to any one of claims 35 to 39, wherein in the ortho position of the phenyl group there is a halo, C alkyl, methoxy or ethoxy susbtituent.

41 . A compound according to any one of claims 35 to 39, wherein in the meta position of the phenyl group there is a C1-4 alkyl or Cs-e heteroayl substituent.

42. A compound according to any one of claims 35 to 39, wherein in the para position of the phenyl group there is an amido or amidomethyl susbtituent.

43. A compound according to claim 38, wherein the phenyl group bears a halo or methoxy substituent in the ortho position, and an amido or amidomethyl substituent in the para position of the phenyl group.

44. A compound according to claim 38, wherein the phenyl group bears an ethoxy substituent in the ortho position, and an amido or amidomethyl substituent in the para position of the phenyl group.

45. A compound according to any one of claims 35 to 39, wherein in the meta position of the phenyl group there is an amino susbtituent.

46. A compound according to any one of claims 1 to 32, wherein A is optionally substituted naphthyl.

47. A compound according to claim 46, wherein A is unsubstituted naphthyl.

48. A compound according to claim 46, wherein A is naphthyl having 1. 2, 3. 4 or 5 substituents.

49. A compound according to claim 46, wherein the naphthyl has 1 or 2 substituents.

50. A compound according to claim 46, wherein the naphthyl has 1 substituent.

51 . A compound according to claim 46, wherein the naphthyl has 2 substituents.

52. A compound according to any one of claims 1 to 32, wherein A is optionally substituted C5-12 heteroaryl.

53. A compound according to claim 52, wherein the C5-12 heteroaryl group is selected from the group consisting of: pyridinyl, pyrimidinyl, pyrazinyl, isoxazoiyi, oxazoiyi, thiophenyl, thiazoiyi, thiadiazoiyi, pyrazoiyi, pyridonyl, imidazoiyi, benzimidazoiyi, imidazopyridinyl and quinolinyl.

54. A compound according to claim 53, wherein the C5-12 heteroaryl group is either pyridinyl or pyrimidinyl.

55. A compound according to claim 53, wherein the C5-12 heteroaryl group is selected from pyridyl, pyrimidinyl, oxazoiyi, oxadiazolyl, pyrazoiyi and thiazoiyi.

56. A compound according to any one of claims 52 to 55, wherein A is unsubstituted C5-12 heteroaryl.

57. A compound according to any one of claims 52 to 55, wherein A is C5-12 heteroaryl having 1. 2, 3, 4 or 5 substituents.

58. A compound according to claim 57, wherein the C5-12 heteroaryl has 1 or 2 substituents.

59. A compound according to claim 58, wherein the C5-12 heteroaryl has 1 substituent.

60. A compound according to claim 59, wherein the C5-12 heteroaryl has 2 substituents.

61 . A compound according to any one of claims 57 to 60, wherein in the ortho position of a Ce heteroayl group, or a-position of C5 and C7-12 heteroaryl group there is a halo or methoxy susbtituent.

62. A compound according to any one of claims 57 to 61 , wherein in the para position of a Ce heteroayl group, or γ-position of Co and CM 2 heteroaryl group there is an amido or amidomethyl.

63. A compound according to any one of claims 57 to 60, wherein in the meta position of a Ce heteroayl group, or in the β-position of a C5 or C7-12 heteroaryl group, there is an amino susbtituent.

64. A compound according to any one of claims 1 to 32, wherein A is selected from one of the following groups:

65. A compound according to any one of claims 1 to 32, wherein A is selected from one of the following groups:

66. A compound according to claim 65, wherein A is selected from one of the following groups:

67. A compound according to any one of claims 1 to 32, wherein A is selected from one of the following groups:

67. A compound according to any one of claims 1 to 32, wherein A is selected from one of the following groups:

68. A compound according to any one of claims 1 to 32, wherein A is selected from one of the following groups: phenyl, with a para-amido substituent and an optional ortho-ethoxy substituent; phenyl, with a para-ether substituent; 6-amino-3-pyridyl; 5-amino-3-pyridyl; and 2-amino-4-pyridyl.

69. A compound according to any one of claims 1 to 68 for use in a method of therapy.

70. A pharmaceutical composition comprising a compound according to any one of claims 1 to 68 and a pharmaceutically acceptable excipient.

71 . A method of treatment of cancer, comprising administering to a patient in need of treatment, a compound according to any one of claims 1 to 68, or a composition according to claim 70.

72. Use of a compound according to any one of claims 1 to 68 in the manufacture of a medicament for treating cancer.

73. A compound according to any one of claims 1 to 68 or pharmaceutical composition according to claim 70, for use in the treatment of cancer.

74. A method of treatment of hemoglobinopathies, comprising administering to a patient in need of treatment, a compound according to any one of claims 1 to 68 or pharmaceutical composition according to claim 70.

75. Use of a compound according to any one of claims 1 to 68 in the manufacture of a medicament for treating hemoglobinopathies.

76. A compound according to any one of claims 1 to 68 or pharmaceutical composition according to claim 70 for use in the treatment of hemoglobinopathies.